Optical Tweezers

EDU-OT2
EDU-OT2/M
Portable Optical Tweezers
User Guide
Portable Optical Tweezers Kit
Table of Contents
Chapter 1 Warning Symbol Definitions ....................................................... 1
Chapter 2 Safety ............................................................................................ 2
Chapter 3 Product Description .................................................................... 3
Chapter 4 Principles of Optical Tweezers ................................................... 5
4.1. Dipole Approach in the Rayleigh Scattering Regime R << ....... 5
4.2. Geometrical Optics Approach in the Mie Regime R >>  ........... 7
Chapter 5 Kit Components ......................................................................... 17
5.1. Trapping Laser Source .............................................................17
5.2. Beam Expander .......................................................................18
5.3. Right-Angle Mirrors ................................................................19
5.4. Sample Positioning System .....................................................20
5.5. Microscope .............................................................................21
5.6. Additional Components...........................................................23
5.7. Screws Included ......................................................................24
Chapter 6 Setup and Adjustment ............................................................... 25
6.1. Assembly ................................................................................25
6.1.1. Trapping Laser Source............................................................................ 26
6.1.2. Beam Expander ...................................................................................... 28
6.1.3. Microscope .............................................................................................. 33
6.1.4. Sample Positioning System .................................................................... 40
6.2. Controller and Software Setup ................................................44

6.2.1. LED Controller ......................................................................................... 44
6.2.2. Laser Controller ....................................................................................... 44
6.2.3. Camera ................................................................................................... 47
6.2.4. Sample Positioning Stages ..................................................................... 50
6.2.5. Kinesis Settings....................................................................................... 53
6.3. Camera Adjustment ................................................................54

6.4. Beam Adjustment ...................................................................55
6.5. Trapping, Microscope Focus, and Final Alignment ...................59
Portable Optical Tweezers Kit
Chapter 7 Experiments ............................................................................... 63

7.1. Creating a Sample .................................................................. 63
7.2. Setting the Correct Focus Level ............................................... 64
7.3. Arranging the Silica Beads ..................................................... 65
7.4. Manipulating a Dairy Cream Particle in a Cream/Water
Emulsion ................................................................................ 66
7.5. The Holding Force of the Optical Trap .................................... 69
7.5.1. Brownian Motion ...................................................................................... 69
7.5.2. The Maximum Holding Force .................................................................. 71
Chapter 8 Teaching Tips ............................................................................ 73

Chapter 9 Control via Game Controller .................................................... 81
Chapter 10 Troubleshooting ........................................................................ 84
Chapter 11 Regulatory .................................................................................. 86
Chapter 12 Thorlabs Worldwide Contacts .................................................. 87
Portable Optical Tweezers Kit Chapter 1: Warning Symbol Definitions
Chapter 1 Warning Symbol Definitions

Below is a list of warning symbols you may encounter in this manual or on your device.
Symbol Description
Direct Current
Alternating Current
Both Direct and Alternating Current
Earth Ground Terminal
Protective Conductor Terminal
Frame or Chassis Terminal
Equipotentiality
On (Supply)
Off (Supply)
In Position of a Bi-Stable Push Control
Out Position of a Bi-Stable Push Control
Caution: Risk of Electric Shock
Caution: Hot Surface
Caution: Risk of Danger
Warning: Laser Radiation
Rev B, July 8, 2019 Page 1

Portable Optical Tweezers Kit Chapter 2: Safety
Chapter 2 Safety
Warning
The class 3B laser diode used in this kit can emit more than 50 mW of optical power,
which can cause damage to the eyes if viewed directly. The laser driver is equipped with
a key switch and safety interlock, which should be used appropriately to avoid injury.
Additionally, we recommend wearing appropriate laser safety glasses
when using this kit.
Page 2 MTN012639-D02
Portable Optical Tweezers Kit Chapter 3: Product Description
Chapter 3 Product Description

For many people, moving and controlling objects with a beam of light sounds more like the
“tractor beams” of science fiction than reality. However, optical tweezers are devices that
allow precisely that kind of manipulation. Many areas of research use them to measure
small forces on the order of piconewtons1. More exotic applications include the control of
tiny microgears2. Biologists use optical tweezers to manipulate different types of molecules
and cells3. In-vitro fertilization of ova is a typical application example – sperm can be
inserted into ova without mechanical contact, thus maintaining a sterile environment.
In a lab course, various demonstrations and experiments can be performed with an optical
tweezers setup. This kit can be used to carry out basic experiments such as moving small
spheres or cells through a solution. The kit can also be used for more advanced
experiments such as investigating the Brownian motion of objects, and measuring the
optical forces of the tweezers.
The working principle can be explained using concepts usually known to undergraduate
students, such as geometric optics, basic theory of Brownian motion, and Stokes’ friction.
It is an intriguing experience to be able to control objects with a laser beam – and not only
for students!
This Optical Tweezers Kit can be assembled into a complete and fully operating
experimental setup with which particles on the order of microns can be trapped and moved.
The beam path is schematically depicted in Figure 1. It is possible to perform a variety of
experiments using a number of different particles such as polystyrene beads, glass beads,
or starch grains from ordinary corn flour. A special feature of this setup is that it is portable.
It can be moved from room to room without needing disassembly or major readjustment,
making it ideally suited to demonstrate the principle of optical tweezers to students in
seminars or lecture halls.
We recommend using the OTKBTK sample kit with the setup. The performance was
optimized for the sample slides and the cover glasses provided with the OTKBTK. For
simplicity, we designed the tweezers system to work without immersion oil.
1
K. SVOBODA, S.M. BLOCK: Optical trapping of metallic Rayleigh particles, Optics Letters 19
(1994) 13, 930-932
2
S.L. NEALE, M.P. MACDONALD, K. DHOLAKIA, T.F. KRAUSS: All-optical control of microfluidic
components using form birefringence, Nature materials 4 (2005), 530-533
3
J.E. MOLLOY, M.J. PADGETT: Lights, action: optical tweezers, Cont. Phys. 43 (2002) 43, 241-
258
Portable Optical Tweezers Kit Chapter 3: Product Description
Camera
Microscope Beam Path
Laser Beam Path
Laser
LED Sample
Position
The light path through the optical tweezers experiment.
Portable Optical Tweezers Kit Chapter 4: Principles of Optical Tweezers
Chapter 4 Principles of Optical Tweezers

To describe the function of optical tweezers, we will examine the force that a focused laser
beam with a Gaussian intensity profile (the TEM00 mode) exerts on an object, which is near
or in the focus. Usually one also assumes that the object is a bead, which consists of a
dielectric, linear, isotropic, and spatially and chronologically non-dispersive material. In the
experiments described below, micron-sized beads made of polystyrene are primarily used.
It is customary to describe the force of the laser on the object by separating it into two
components. One component, the scattering force, acts along the direction of beam
propagation. The second component acts along the intensity gradient and is therefore
called the gradient force. The gradient force can act in different directions with respect to
the beam. As the laser has a Gaussian intensity profile, the gradient force can act
orthogonally to the beam, but it can also act parallel to the beam, as the laser is focused
and therefore also has an intensity gradient along the beam axis. These two components
and their relationship to one another are the defining factors for whether or not a particle
can be trapped by the optical trap. Stable optical tweezers are only obtained if the gradient
force, which pulls the object in the direction of the focus, is greater than the scattering
force, which pushes the particle in the direction of the beam away from the focus.
The various theoretical approaches to describe optical trapping can roughly be divided
according to the areas in which they are valid. The relationship of the radius 𝑅 (or diameter
𝑑) of the bead to the wavelength 𝜆 of the incident laser beam is the dividing factor. The
case 𝑅 ≈ 𝜆 is theoretically very complex and shall therefore not be dealt with here. The
two extreme cases for very large and very small particles are summarized below:
4.1. Dipole Approach in the Rayleigh Scattering Regime R << 

The first case we will consider is when the radius 𝑅 of the bead is significantly smaller than
⃗ (𝑟) is approximately
the wavelength 𝜆 of the incident laser beam. Then, the electrical field 𝐸
spatially constant with respect to the particle and the situation can be portrayed as follows:
As the bead is assumed to be dielectric, one can imagine it as a collection of 𝑁 point
dipoles. Due to their polarizability, a dipole moment 𝑝𝑖 is induced in each of the point
dipoles by the incident laser beam. Due to the linearity of the material, the following applies:
⃗ (𝑟𝑖 )
𝑝𝑖 = 𝛼 ⋅ 𝐸 (1)
Here, 𝑟𝑖 is the location of the i-th point dipoles and 𝐸 ⃗ (𝑟𝑖 ) is the electrical field strength at
this location. In addition, the electrical field of the laser appears to be approximately
spatially constant for the bead due to the condition 𝑅 ≪ 𝜆, meaning that at a certain point
in time 𝑡0 the strength of the electrical field is equally great for all point dipoles of the bead.
As a result, the induced dipole moment is equally great for all 𝑁 point dipoles. The
polarization 𝑃 resulting from the induced dipole moments is then

1 𝑁
𝑃 = ∑𝑝 = ⃗ = 𝜒 ⋅ 𝜖0 ⋅ 𝐸⃗
⋅𝛼⋅𝐸 (2)
𝑉 𝑉
𝑖
where 𝜒 is the electrical susceptibility, 𝜖0 is the electrical constant, and 𝑉 is the volume of
the bead. The potential energy 𝑈𝑖 of one of the point dipoles with dipole moment 𝑝 in the
⃗ is 𝑈𝑖 = −𝑝𝐸⃗ . Because there are 𝑁 point dipoles in a bead with the volume
electrical field 𝐸
𝑉, the energy density in the bead is defined by:
𝑁 ⋅ 𝑈𝑖 𝑁
𝑈= = − ⋅ 𝑝 ⋅ 𝐸⃗ = −𝑃⃗ ⋅ 𝐸⃗ (3)
𝑉 ⏟
𝑉
𝑃⃗
The occurrence of the gradient force, which is a force component that is directed in the
direction of the intensity gradient of the incident electrical field, can be explained when one
observes this potential energy 𝑈 of the bead in the electrical field. Equation (2) states that
2
𝑃 is proportional to 𝐸⃗ . Therefore, according to equation (3), 𝑈 is proportional to |𝐸⃗ | and
2
thus to intensity 𝐼 ∝ |𝐸⃗ | of the incident field. The force exerted on the particle by the
incident field is proportional to the gradient of the potential energy ∇𝑈 and therefore
proportional to the intensity gradient ∇𝐼. The following equations describe the gradient
force:
2𝜋𝛼
𝐹𝐺𝑟𝑎𝑑 = ∇𝐼 (4)
𝑐 𝑛2𝑚
2
𝑚2 − 1
𝛼 = 𝑛𝑚 𝑅3 ( ) (5)
𝑚2 + 2
𝑛𝑝
𝑚= (6)
𝑛𝑚
Here, 𝛼 is the polarizability of the dipoles and 𝑚 is the relationship of the refraction index
of the particles, 𝑛𝑝 (polystyrene in our case) to the refraction index of the surrounding
medium, 𝑛𝑚 (water in our case).
The destabilizing scattering force component is explained by the scattering of the incident
light at the particle. The force action is created by the absorption and isotropic re-emission
of the light by the bead. As 𝑅 ≪ 𝜆, the conditions are fulfilled for Rayleigh scattering. The
resulting force can be stated as follows:
𝜎 𝑛𝑚
𝐹𝑆𝑐𝑎𝑡𝑡𝑒𝑟𝑖𝑛𝑔 = ⋅ 𝐼 (7)
𝑐
2
128 𝜋 5 𝑅6 𝑚2 − 1
𝜎 = ( 2 ) (8)
3 𝜆4 𝑚 +2
Here, 𝑐 is the speed of light in vacuum and 𝐼 is the incident intensity and 𝜎 is the scattering
cross-section of the incident light. It is important to note that the scattering force is
proportional to the intensity and points in the beam direction.
4.2. Geometrical Optics Approach in the Mie Regime R >> 

This ray optical approach deals with the second possible extreme case. Here, we will
assume that the radius of the bead is much larger than the wavelength of the incident
laser. In this range, the conditions of geometric (ray) optics are fulfilled and one can think
of the laser beam as a bundle of rays. Typically, this assumption is valid for beads with a
radius 𝑅 that is greater than ∼ 10𝜆. The basics of the theoretical derivation of the gradient
and scattering force in accordance with this model can be found in a work by Ashkin4.
The particle properties of light must now be taken into account, namely that light can
transfer momentum to an object in the form of photons. The force action of a beam on a
particle can be explained using Newton’s second law: the force on a particle is exactly
equal to the change in the momentum of the particle over time:
𝑑𝑝
𝐹= (9)
𝑑𝑡
𝑑𝑝
The following equation describes the change in momentum of a beam over time in a
𝑑𝑡
medium with a refractive index 𝑛𝑚 :
𝑑𝑝 𝑛𝑚 𝑛𝑚
= 𝑃𝐵𝑒𝑎𝑚 = 𝐼(𝑟)𝑑𝐴 (10)
𝑑𝑡 𝑐 𝑐
Here, 𝐼(𝑟) is the intensity distribution in the beam cross-section. Often, a Gaussian profile
is used, in which the intensity decreases in a Gaussian distribution from the center of the
beam outward. This is also the case in our setup.
If a beam with the power 𝑃𝐵𝑒𝑎𝑚 hits a sphere at an angle of 𝜗, part of the beam will be
reflected and part of it will reach the interior of the sphere through transmission (see Figure
2). For the power of these two partial beams, the following is in effect:
𝑃𝑟𝑒𝑓𝑙 = 𝑃𝐵𝑒𝑎𝑚 ⋅ 𝑅𝑟 (11)
𝑃𝑡𝑟𝑎𝑛𝑠 = 𝑃𝐵𝑒𝑎𝑚 ⋅ 𝑇 (12)
Here, 𝑅𝑟 is the reflectivity and 𝑇 is the transmissivity. The transmitted beam transports
momentum into the sphere in accordance with the equation (10).
4Ashkin A., Forces of a single-beam gradient laser trap on a dielectric sphere in the ray
optics regime. In: Biophys. J. 61 (1992) 2, 569-582
Reflection and Transmission of an Incident Partial Beam on the

Inside and Outside Surfaces of a Sample Bead with a Refractive Index
Higher than the Immersion Medium
Inside the sphere, the beam is reflected and transmitted numerous times. Part of the beam
is repeatedly reflected on the sphere's internal wall and remains in the sphere, while the
rest exits the sphere again through transmission. The beams which exit the sphere again
𝑑𝑝
were thus subject to a change in momentum . The force on the sphere is equal to the
𝑑𝑡
momentum per unit time that remains in the sphere, based on equation (9). Now, the force
on the sphere is once again divided into two components: a component in the direction of
the incident beam (corresponds to the Z direction) and a perpendicular component
(corresponds to the Y axis). This results in the following for both forces:
𝑃𝐵𝑒𝑎𝑚
𝐹𝑠 ≔ 𝐹𝑧 = 𝑛𝑚 𝑄𝑠 (13)
𝑐
with the Q factor
𝑇2 (cos(2𝜃 − 2𝑡) + 𝑅𝑟 cos(2𝜃))

𝑄𝑠 = 1 + 𝑅𝑟 cos(2𝜃) − (14)
1 + 𝑅2𝑟 + 2𝑅𝑟 cos(2𝑡)
and
𝐹𝑔  𝐹𝑦 = 𝑛𝑚 𝑄𝑔 (15)
𝑐
with the Q factor
𝑇2 (sin(2𝜃 − 2𝑡) + 𝑅𝑟 cos(2𝜃))

𝑄𝑔 = 𝑅𝑟 sin(2𝜃) − (16)
1 + 𝑅2𝑟 + 2𝑅𝑟 cos(2𝑡)
Here, 𝑡 is the angle at which the first transmitted beam is refracted toward the normal (see
Figure 2). According to Snell’s law of refraction, the following relationship is in effect for
the angles 𝜃 and 𝑡:
sin(𝜃) 𝑛𝑝
= (17)
sin(𝑡) 𝑛𝑚
𝑄𝑠 and 𝑄𝑔 are dimensionless Q factors, which state what percentage of the incident
momentum contributes to the force parallel or perpendicular to the beam, respectively.
These factors depend heavily on the angle of incidence of the beam, as one can see from
the equations. This angle becomes larger the more heavily the beam is focused, which
occurs when a higher numerical aperture objective is used.
The component of the beam that points in the incident direction (Z direction) ultimately
causes the scattering force 𝐹𝑠 . The component perpendicular to this (Y direction) is mainly
responsible for the gradient force 𝐹𝑔 . In order to obtain the overall power, one must
naturally consider all partial beams and integrate all of them. That will be discussed in
detail below.
Q Factor Angular Dependence5
5 Ashkin A., Forces of a single-beam gradient laser trap on a dielectric sphere in the ray optics regime.
In: Biophys. J. 61 (1992) 2, 569-582
Figure 3 shows the values of the two 𝑄 −factors, depending upon the angle of incidence
𝜃 when the focus is located slightly above the surface of the sphere. One can see here 𝑄𝑔
is negative through almost the entire range, meaning the force acts in the negative Y
direction, which is upwards in Figure 2. The factor 𝑄𝑠 is always positive. The Z-component
of the force therefore always points in the beam direction. If the beam were not to hit in the
upper half of the sphere, but rather in the lower half, one can easily conclude for reasons
of symmetry that the direction of the 𝑦 −component would reverse, the direction of the Z-
component would remain the same.
In short: The sphere always moves into the focus or the point of highest intensity. In order
to ultimately achieve a stable optical trap, the following must be true:
𝐹𝑔 > 𝐹 𝑠 (18)
In the following, we’ll discuss these forces in more detail.
Total force on the sphere

To obtain the total force acting on the sphere, we have to sum over all partial beams that
hit it.
For that, we integrate (i) over the distance 𝑟

between the partial beam and the symmetry
axis of the whole beam, ranging from 0 to
𝑟𝑚𝑎𝑥 , and (ii) over its angular coordinate 𝛽,
ranging from 0 to 2.
Figure 4 shows how the coordinates 𝑟 and 𝛽

of a partial beam (in red) are defined. The
dashed line stands symbolically for the
sphere the laser is focused on.
Coordinates of a Partial Beam
Page 10 MTN012639-D02
So far, the force was given as a function of 𝜃 which now has to be expressed in terms of
𝑟 and 𝛽. Figure 5 shows a partial beam incident on the sphere with angle 𝜃 from the side.
The following statements hold true:
𝑋 = sin 𝜙 ∙ 𝑆 = sin 𝜃 ∙ 𝑅 (19)

and
(20)
sin 𝜙 = 𝑟⁄√𝑟 2 + 𝑙2
𝑙 roughly corresponds to the focal length of the objective. Effectively, you can use the
objective’s working distance for this parameter.
Relation Between 𝒓 and 𝜽

Then, 𝜃 can be expressed as
𝑆 𝑟
𝜃(𝑟) = arcsin ( ) (21)
𝑅 √𝑟 2 + 𝑙 2

Next, the forces need to be summed. For that, we start by observing Figure 6. A partial
beam with distance 𝑟 to the symmetry axis of the whole beam falls on the sphere under
an angle 𝜃. As discussed above, we can split the resulting force in two perpendicular
components, 𝐹𝑠 and 𝐹𝑔 . For clarity, we can now add another partial beam, namely the one
mirrored on the symmetry axis, denoted “mirror beam”. As sketched in Figure 6, this partial
beam falls on the sphere on the other side and results in “mirrored” force vectors 𝐹𝑠,𝑚𝑖𝑟𝑟𝑜𝑟
and 𝐹𝑔,𝑚𝑖𝑟𝑟𝑜𝑟 in the right part of the sphere (which were not drawn to avoid an overcrowded
figure).
The Contribution of One Ray to the Total Force5,6

When you consider all of the partial beams with the same condition around the symmetry
axis, it immediately becomes clear that all force components in X and Y direction vanish
and only resulting force components along the Z axis remain.
These can be written as
𝑟
𝐹𝑔,𝑧 = −𝐹𝑔 ∙ sin 𝜙 = −𝐹𝑔 ∙ (22)
√𝑟 2 + 𝑙 2
𝑙
𝐹𝑠,𝑧 = 𝐹𝑠 ∙ cos 𝜙 = 𝐹𝑠 ∙ (23)
√𝑟 2 + 𝑙 2
where 𝐹𝑔,𝑧 is pointing in the negative Z direction and is, therefore, negative. Hence, each
infinitesimal force contributing to the total force is given by
𝑑𝐹 = 𝐹𝑠 ∙ cos 𝜙 − 𝐹𝑔 ∙ sin 𝜙 (24)
6
Adapted from A. Langendörfer:”Aufbau einer Optischen Pinzette für das Landesmuseum für
Technik und Arbeit in Mannheim“, wissenshaftliche Arbeit, KIT, Karlsruhe, 2009
Page 12 MTN012639-D02
The total force 𝐹𝑡𝑜𝑡 is the sum of the total scattering force 𝐹𝑠,𝑡𝑜𝑡 and the total gradient force
𝐹𝑔,𝑡𝑜𝑡 , where
𝑟𝑚𝑎𝑥 2𝜋
𝐹𝑠,𝑡𝑜𝑡 = ∫ ∫ 𝐹𝑠 ∙ cos 𝜙𝑟𝑑𝑟𝑑𝛽 (25)

0 0
and
𝑟𝑚𝑎𝑥 2𝜋
𝐹𝑔,𝑡𝑜𝑡 = ∫ ∫ −𝐹𝑔 ∙ sin 𝜙𝑟𝑑𝑟𝑑𝛽 (26)

0 0
In Figure 7, we plotted the gradient force, the scattering force and the resulting total force
as a function of the focus position 𝑆 relative to the radius 𝑅 of the sphere. The direction is
the Z axis, meaning that the plot shows the behavior when the focus is moved through the
center of the sphere from bottom to top. The curves were calculated with the above
equations with a set of typical parameters since we only want to discuss the curve’s
general form. Therefore, the forces are given in arbitrary units (a.u.). On the horizontal
axis, “+1” corresponds to the focus position on the sphere’s outer surface right above the
sphere’s center while “-1” corresponds to the lower surface below the center.
Gradient force, 𝐹𝑔,𝑡𝑜𝑡

Scattering force, 𝐹𝑠,𝑡𝑜𝑡
Total force, 𝐹𝑡𝑜𝑡
Force [a.u.]
Location of the focus in units of the sphere radius 𝑅 (Z direction)

Overview of All Forces When Focus is Changed in Z-Direction
From Figure 7, we learn that
1. The gradient force always points towards the focus. For example, when 𝑆/𝑅 ≥ 0,
the focus is above the sphere’s center. Then 𝐹𝑔 ≤ 0 and the gradient force pushes
the sphere upwards into the focus.
2. The scattering force always points in the direction of propagation of the incident
beam, in this case downwards. Note that 𝐹𝑠 ≥ 0 in the entire range of the plot.

3. When the focus is below the sphere’s center, i.e. 𝑆/𝑅 ≤ 0, scattering and gradient
force act in the same direction. When the focus is above the sphere’s center,
i.e. 𝑆/𝑅 ≥ 0, both forces point in opposite directions.
We also note that the absolute value of the gradient force is always higher than the
absolute value of the scattering force. This is the requirement for any stable optical
tweezers trap. The fundamental quantity to fulfill this requirement is the microscope
objective’s numerical aperture, which defines the angle of the focus which we will discuss
next.
Sometimes the focus is not above or below the sphere’s center but along the y-axis
instead. Again, the reference axis goes through the sphere’s center, and we plot the
occurring forces as a function of the focus’ distance 𝑆 to the sphere’s center in units of the
sphere’s radius.
Figure 8 shows the gradient force and the scattering force. Plotting the sum of both forces
would not make sense as they point in different directions (𝐹𝑠,𝑡𝑜𝑡 in Z direction and 𝐹𝑔,𝑡𝑜𝑡 in
Y direction).
From Figure 8, we learn that:
1. The general form of the curves is similar to the focus’ movement on the Z axis.
Only outside of the sphere the force decreases a little faster than compared to
the Z axis.
2. The maximal gradient force is larger than the maximal scattering force (absolute
values). This is the case when the parameters such as the numerical aperture
allow optical trapping.
3. The maximal gradient force are located just within the sphere, close to the
surface. The maximal value of the scattering force is found right on the surface.
Gradient force, 𝐹𝑔,𝑡𝑜𝑡

Scattering force, 𝐹𝑠,𝑡𝑜𝑡
Force [a.u.]
Location of the focus in units of the sphere radius 𝑅 (Y direction)
Scattering and Gradient Force with the Focus Changed in Y

Direction6
Page 14 MTN012639-D02
Influence of the Numerical Aperture

As we have seen above, the angle of incidence of the partial rays plays a crucial role in
optical tweezing. The angle is defined by the numerical aperture of the objective: the
numerical aperture (𝑁𝐴) describes the acceptance cone of an objective and is given by
𝑁𝐴 = 𝑛 ∙ sin 𝜙 (27)
where 𝑛 is the refractive index of the material between the objective and the focus and 𝜙
is half of the angle of the maximum light cone.
As discussed, the gradient force 𝐹𝑔,𝑡𝑜𝑡 needs to exceed the scattering force 𝐹𝑠,𝑡𝑜𝑡 to get a
stable trap. Next, we want to investigate a measure for the trap’s strength. For that, we
can have a look at the ratio of 𝐹𝑔,𝑡𝑜𝑡 and 𝐹𝑠,𝑡𝑜𝑡 at the point 𝑆/𝑅 = 1 since we have shown
that scattering and gradient force are strongest when the laser focus is at/near the sphere’s
surface (i.e., |𝑆⁄𝑅 | = 1). So for a stable trap, we can assume the condition:
𝐹𝑔,𝑔𝑒𝑠 (𝑆⁄𝑅 = 1)
| |≥1 (28)
𝐹𝑠,𝑔𝑒𝑠 (𝑆⁄𝑅 = 1)
Absolute values are used since 𝐹𝑔,𝑡𝑜𝑡 is negative. Figure 9 shows how this strength
depends on the numerical aperture. Again, we plotted a curve with typical parameters to
show the general behavior of the curve; therefore, we do not focus on the concrete
numbers.
Figure 9 shows the fundamental behavior that the strength of the trap increases with
increasing numerical aperture. Also, it becomes apparent that there is a lower limit for the
numerical aperture of the objective. Below that, no trapping occurs since the gradient force
never exceeds the scattering force.
|𝐹𝑔,𝑡𝑜𝑡 / 𝐹𝑠,𝑡𝑜𝑡 |
Numerical aperture 𝑁𝐴
Behavior of the Trap’s Strength with Respect to the Numerical

Aperture6

Influence of the Laser Power

While the numerical aperture affects the ratio of gradient and scattering force, the laser
power directly influences the individual forces. In Equations (13)
𝐹𝑠 ≔ 𝐹𝑧 = 𝑛𝑚 𝑄𝑠
𝑐
and (15),
𝐹 𝑔  𝐹 𝑦 = 𝑛𝑚 𝑄𝑔
𝑐
we see that both forces increase proportionally with the laser power 𝑃. When the laser
power is increased, each individual force gets stronger. However, their ratio (which
essentially determines if optical tweezing ocurrs or not) remains the same, see Equation
(28).
Combining what we know about the numerical aperture and laser power, we can note that
the increase in trapping strength can be achieved through two factors:
 Increase of the numerical aperture of the objective so that the gradient force
exceeds the scattering force.
 Increase of the laser power, which increases the tweezer’s strength linearly if the
condition 𝐹𝑔,𝑡𝑜𝑡 > 𝐹𝑠,𝑡𝑜𝑡 is met.
This also tells us that increasing the numerical aperture is the more effective way to
improve trapping than increasing the laser power. While the laser power increases the
trapping force linearly, Figure 9 shows a stronger than linear behavior above the trapping
limit.
Page 16 MTN012639-D02
Portable Optical Tweezers Kit Chapter 5: Kit Components
Chapter 5 Kit Components

In cases where the metric and imperial kits contain parts with different item numbers,
metric part numbers and measurements are indicated by parentheses unless otherwise
noted.
5.1. Trapping Laser Source
1 x SR9A-DB9 1 x LTN330-A
ESD Protection and Strain 1 x L658P040 Adjustable Collimator for
Relief Cable 658 nm, 40 mW, Ø5.6 mm Laser Diodes,
Ø5.6 mm, A Pin Code AR Coated: 350 – 700 nm
Laser Diode
1 x TPS002
±15 V / 5 V K-Cube
1 x KLD101 Power Supply 1 x RS3.5P8E
K-Cube Laser Diode (RS3.5P4M)
Driver Ø1" (Ø25 mm) Pedestal
Post, 3.5" (90 mm) Tall
1 x AD15F
1 x CF125 SM1-Threaded Adapter for
1 x KC1-T(/M)
Small Clamping Fork Ø15 mm Components
Ø1" Cage-Compatible
SM1-Threaded Mirror
Mount

5.2. Beam Expander
2 x ER1 2 x ER3
2 x ER10
Ø6 mm Cage Assembly Ø6 mm Cage Assembly
Ø6 mm Cage Assembly
Rod, 1" Long Rod, 3" Long
Rod, 10" Long
2 x ER6
Ø6 mm Cage Assembly 4 x CP09(/M) 1 x LA1074-A
Rod, 6" Long 30 mm Removable Ø1/2" N-BK7 Plano-
Segment Cage Plate Convex Lens, f = 20 mm,
AR Coating: 350-700 nm
1 x LA1509-A 1 x SM1A6 1 x SM05L03

Ø1" N-BK7 Plano-Convex Adapter with External Ø1/2" Lens Tube,
Lens, f = 100 mm SM1 Threads and Internal 0.3" Long
AR Coating: 350-700 nm SM05 Threads
1 x CF125
1 x TR3 (TR75/M) Small Clamping Fork
1 x PH3 (PH75/M)
Ø1/2" (Ø12.7 mm) Post,
Ø1/2" (Ø12.7 mm) Post
3" (75 mm) Long
Holder, 3" (75 mm) Long
1 x BE1(/M)
Pedestal Base Adapter
Page 18 MTN012639-D02
5.3. Right-Angle Mirrors
2 x PF10-03-P01 2 x SM1L05
2 x KCB1(/M) Ø1" Protected Silver Ø1" Lens Tube, 0.5"
Right-Angle Kinematic Mirror Long
Mirror Mount, 30 mm
Cage Compatible
2 x SM1L10 2 x SM1CPL10
Ø1" Lens Tube, 1" long SM1 Lens Tube Flexure
Sleeve Coupler, 1" Long

5.4. Sample Positioning System
2 x MT1(/M)-Z8
1 x MT1B(/M)
Motorized Translation
Manual Translation Stage, 2 x KDC101
Stage, 1/2" (12 mm)
1/2" (13 mm) Travel7 K-Cube DC Servo Motor
Travel
Module
1 x MT401(/M)
Mounting Base for
1 x MT402
Translation Stages
2 x KPS101 Right Angle Bracket
K-Cube Power Supply
2 x MT405 1 x Slide Holder Part 1 1 x Slide Holder Part 2

Side-Mounted Actuator
Adapter for MT1(/M)-Z8
7
The screw of the MT1B(/M) allows for a travel of 150 µm per revolution, which is much finer than
the 500 µm per revolution provided by a similarly-sized micrometer.
Page 20 MTN012639-D02
5.5. Microscope
1 x DCC1645C 1 x SM1L15
High-Resolution CMOS 2 x SM1T2 Ø1" Lens Tube, 1.5" Long
Camera Ø1" Lens Tube Coupler
2 x FGB37
Ø25 mm BG40 Colored
Glass Bandpass Filter, 1 x SPT1(/M) 4 x CP02(/M)
335 – 610 nm Coarse ±1 mm XY Slip SM1-Threaded Cage
Plate Positioner Plate
1 x LB1676
3 x C1498(/M) Ø1" N-BK7 Bi-Convex 1 x CP02T(/M)
Slip-On Ø1.5" Post Lens, f = 100 mm SM1-Threaded Cage
Clamps Plate, 0.5" Thick
1 x FES0650
Ø1" Shortpass Filter, 1 x CCM1-4ER(/M) 1 x BS022
650 nm Cut-Off Empty 30 mm Cage Cube, 70:30 (R:T) Non-
8-32 (M4) Tap Polarizing Beamsplitter
Cube, 400 - 700 nm, 1"

1 x SM1A17
1 x SM1CP2 1 x Zeiss Microscope
Adapter with External
SM1 End Cap Objective
SM1 Threads and Internal
M27 x 0.75 Threads 63X, 0.8 NA
4 x ER05
Ø6 mm Cage Assembly 4 x ER3 4 x ER1.5
Rod, 0.5" Long Ø6 mm Cage Assembly Ø6 mm Cage Assembly
Rod, 3" Long Rod, 1.5" Long
1 x DG10-600
1 x MCWHL5 Ø1" N-BK7 Ground Glass
1 x SM1L10 Diffuser, 600 Grit
Cold White Mounted LED Ø1" Lens Tube, 1" Long
1 x LEDD1B
T-Cube LED Driver 1 x KPS101
Power Supply 1 x DP14A(/M)
Damped Ø1.5" Post,
14" Long
Page 22 MTN012639-D02
5.6. Additional Components
1 x MB1224 (MB3060/M) 1 x RDF1

Aluminum Breadboard, Rubber Dampening 1 x CPA1
12" x 24" x 1/2" Breadboard Feet 30 mm Cage System
(300 mm x 600 mm x 12.7 mm) Alignment Plate with
Ø1 mm Aperture
1 x BBH1
Breadboard Handles 1 x SPW606
(Pack of 2) SM1 Spanner Wrench,
1" Long

5.7. Screws Included

Imperial Kit
Type Qty. Type Qty.

1/4"-20 Cap Screw, 0.315" Long 2 8-32 Cap Screw, 3/8" Long 2
1/4"-20 Cap Screw, 3/8" Long 10 8-32 Cap Screw, 1/2" Long 3
1/4"-20 Cap Screw, 1/2" Long 12 #8 Washer 2
1/4"-20 Cap Screw, 5/8" Long 8 8-32 Setscrew, 1/2" Long 1
1/4"-20 Cap Screw, 3/4" Long 4 1/8" x 1/4" Steel Dowel Pin 8
SD1 8-32 to 1/4"
1/4" Washer 22 1
Counterbore Adapter, 10-Pack
1 x CCHK
Imperial Hex
1 x BD-3/16L Key Set
3/16" Balldriver
Metric Kit
Type Qty. Type Qty.

M6 x 8 mm Cap Screw 2 M4 x 10 mm Cap Screw 2
M6 x 10 mm Cap Screw 10 M4 x 12 mm Cap Screw 3
M6 x 12 mm Cap Screw 12 M4 Washer 2
M6 x 16 mm Cap Screw 8 M4 x 12 mm Set Screw 1
M6 x 20 mm Cap Screw 4 1/8" x 1/4" Steel Dowel Pin 8
SD1 M6 to M4
M6 Washer 22 1
Counterbore Adapter, 10-Pack
1 x CCHK/M
Metric Hex Key
1 x BD-5ML Set
M6 Balldriver
Page 24 MTN012639-D02
Portable Optical Tweezers Kit Chapter 6: Setup and Adjustment
Chapter 6 Setup and Adjustment

6.1. Assembly
The tweezers will be set up on an MB1224 (MB3060/M) breadboard. Set everything up on
a stable desk or table without any sources of vibration nearby. Further damping, such as
using an isolated optical table, is not necessary. First, unpack the balldriver BD-3/16L (BD-
5ML), the SM1 spanner wrench SPW606, and the hex keys CCHK(/M).
Figure 10 shows the breadboard, and the positions of components to be mounted are
indicated with cap screws and red outlines. Attach the BBH1 breadboard handles to the
edges of the breadboard using four 1/4"-20, 5/8" long (M6 x 16 mm) cap screws with
washers so the assembled tweezers can be moved. Attach the RDF1 rubber breadboard
feet to the underside of the breadboard at the positions indicated by the red stars in the
diagram below using four 1/4"-20, 1/2" long (M6 x 12 mm) cap screws without washers.
Use the threaded hole in the breadboard and not the counterbored holes.
Clamping Forks
Damping Post
Breadboard
Handle
Sample
Positioning
Stage
Sample Stage Controllers Laser Driver
LED
Controller
Positions of Components on the Breadboard

Teaching Note:
In the previous version of this kit, EDU-OT1(/M), the laser segment had to be assembled
first and the microscope section built step by step by attaching components to the laser
segment. We received feedback that some customers prefer to build the microscope
section first to demonstrate the fact that optical tweezers can be built by coupling a laser
into an existing microscope.
For this kit, we changed some parts so the microscope and the laser assembly can be built
separate from each other. The laser assembly now features variable tube elements that
allow for a retroactive height match between the two assemblies. Therefore, you may
choose your preferred way of teaching & assembling this kit. Either build the microscope
first and attach the cage segment/laser source to it or the other way around. In this manual,
we present the instructions on how to assemble the laser source and cage segment first.

6.1.1. Trapping Laser Source

Figure 11 shows the components that are necessary for setting up the laser source.
Laser Source Components
First, screw the 8-32, 1/2" long (M4 x 12 mm) setscrew into the
RS3.5P8E (RS3.5P4M). Screw on the KC1-T(/M) in the orientation
depicted to the right (note the position of the tip/tilt adjusters). This
orientation is critical because you will not be able to access the
cage rod locking screws if the KC1-T(/M) is in a different
orientation.
Locking Screws
Next, remove one retaining ring from the mount. Screw in the AD15F
until the threaded region is within the mount and its locking screws
face upwards. Tighten the KC1-T’s remaining retaining ring against
the AD15F using the SPW606. Now feed the SR9A-DB9 through the
mount as shown below.
Page 26 MTN012639-D02
Note: We recommend using a grounding bracelet when

handling the laser diode to prevent damage.
Take the L658P040 laser diode from its package and place
it in the silver retention ring of the LTN330-A, see the image
to the right.
Remove the retaining ring from the LTN330-A housing.

Then, insert the diode with the silver retention ring into the
LTN330-A.
Fix the assembly in the LTN330-A using the black retaining

ring (the retaining ring can be tightened using a thin
screwdriver). Make sure to use the thick retaining ring and
not the one you unscrewed in the previous step. Also,
ensure that the laser diode’s legs fit through the center hole
so they are not damaged, see image to the right.
In the next step, insert the laser diode’s legs into the socket
of the SR9A-DB9. Please be careful in this step to avoid
damage to the diode.
When the diode’s legs are in the socket, the cover can be screwed onto the LTN330-A.
Make sure to screw on the part labeled “SR9A-DB9” instead of rotating the part with the
printed circuit board!
Finally, secure the

assembly in the AD15F by
engaging the two nylon-
tipped setscrews, see
image to the right.

6.1.2. Beam Expander

In the following, the cage system is set up to house the beam expander. The individual
components are assembled first, and then mounted in the cage system.
Adjustment Mirror
Mount the mirrors (PF10-03-P01) in the right angle mirror mounts (KCB1(/M)) with the
reflective surface facing away from the knobs (Figure 12).
Mirror and Right Angle Mount

Base
Screw the BE1(/M) base into the PH3 (PH75/M) post holder as shown in Figure 13.
Post Holder Base

Lens 1
The elements that are necessary for the setup of lens (1), are shown in Figure 14. First,
place the LA1074-A lens into the SM05L03 Ø1/2" lens tube using the included SM05
retaining ring. Ensure that the convex side of the lens is inserted into the lens tube first.
Then, screw the lens tube into the SM1A6 adapter, and attach it to the CP09(/M) cage
plate.
Beam Expander Lens 1 Components

Page 28 MTN012639-D02
Lens 2
The components for lens 2 are shown in Figure 15. The lens (LA1509-A) must be attached
to the CP09(/M) cage plate using two SM1 retaining rings.
Beam Expander Lens 2 Components
Cage Assembly
The cage assembly features a removable segment that comprises the two lenses for beam
expansion. Step by step assembly instructions follow below. Figure 16 demonstrates the
working principle of the removable cage plate assembly.
Function of the Removable Cage Segment

To assemble the cage segment, first screw an empty CP09(/M) onto the
TR3 (TR75/M) in the orientation shown in the image to the right and insert it
into the post holder assembled earlier.
Next, loosen the locking screws on the two lower holes of the CP09(/M),
feed the two ER10 rods through, and screw them into the lower holes of the
KCB1(/M). Screw the two ER1 rods into the two upper holes and remove
the small setscrews at the other side. Move the CP09(/M) with respect to
the cage rods until the ER1 fill half of the CP09(/M)’s flexure locks. Tighten
the side-located 4-40 setscrews (the small/lower ones!) on the CP09(/M)
using a 0.050" (1.3 mm) hex key. The assembly should look like the image
below.
To assemble the beam expander segment, take the two lenses (1) and (2), see above,
and feed the ER6 rods through their cage holes as shown in the photo below. Remove the
cage rods’ setscrews. The distance between the lens centers should be about 12 cm.
Lens Orientation
Next, click the beam expander segment into the assembly. Note that the 4-40 (M3) cap
screws of the CP09(/M)’s flexure mechanism should not be tightened. Take the remaining
CP09(/M) cage plate and slip it onto the assembly. Again, the cage rods should only fill
half of the flexure locks.
Page 30 MTN012639-D02
Fourth CP09(/M)
Cage Plate
Added Last
Finally, attach the kinematic mount with the laser source from Section 6.1.1. To do this,
feed the ER10 cage rods through the lower holes of the kinematic mount (you may need
to loosen the KC1-T’s rod locking screws first). Insert the ER3 cage rods (with removed
setscrews) into the upper holes so that they fill the rest of the flexure locks. You may want
to leave a small gap between the adjacent cage rods. Fix all of the KC1-T’s cage rod
locking screws. You may fix the CP09(/M)’s flexure locks by tightening the 4-40 (M3) cap
screws with a 3/32" (2.5 mm) hex key. However, in our tests, the flexure locks have
demonstrated a sufficient clamping force without tightening the flexure mechanism (which
makes taking the expander section out easier).
Use a level to adjust the height of the post in its post holder. The cage segment
needs to be horizontal to avoid serious problems during alignment/adjustment!
Fully assembled cage segment

Mirror Position
The mirrors are connected to each other and to the microscope with the help of a lens tube
flexure sleeve coupler, SM1CPL10. This allows for freedom in height and distance
adjustment when connecting the laser/cage assembly to the microscope assembly.
For the vertical connection, please use SM1L05 lens tubes. For the horizontal connection,
please use the longer SM1L10 tubes. This ensures that there is sufficient space for manual
adjustment between the stage assembly and the mirrors. Remove the retaining rings from
all four lens tubes.
Screw an SM1L05 lens tube into the KCB1(/M) of the assembly above. Take the second
KCB1(/M) and screw in another SM1L05. Connect them with an SM1CPL10 by slipping it
over the tubes and tightening its locking screws. For them to stay parallel, you can press
a flat object like a book or level on their side.
Screw an SM1L10 lens tube to the free side of the upper KCB1(/M). Slip the second
SM1CP10 on it. Later, that will be connected to a lens tube attached to the beamsplitter.
SM1CPL10
Lens Tube
Couplers
Laser & Cage Segment Fully Assembled
Page 32 MTN012639-D02
6.1.3. Microscope
Attach the damped DP14A(/M) Ø1.5" mounting post to the breadboard using the mounting
holes in its base and four 1/4"-20, 3/4" long (M6 x 20 mm) cap screws. Choose the position
shown in Figure 10. In the following procedure, the individual parts of the microscope must
be built before the microscope system can be created.
Beamsplitter
The BS022 beamsplitter has to be installed in the CCM1-4ER(/M) housing. To do so,
remove the small black screws on the side of the housing with the included hex key and
pull off the top of the housing. Now unscrew the silver screws on the inner section and take
the beamsplitter out of its packaging.
Place the inner piece of the housing on the table in the orientation shown in Figure 19, with
the triangular section at the bottom left side. Also, orient the beamsplitter cube as it is
depicted in Figure 19. In particular, this means that the labels are not visible (“BS022” and
three arrows) and the beamsplitting surface goes from bottom left to top right. Install the
beamsplitter into the inner housing piece in that orientation and tighten the silver screws
so that the triangular upper section secures the cube.
Cover the piece with the outer part of the CCM1-4ER(/M) in the orientation shown in Figure
19 and fix it with the black screws. Finally, screw in four ER05 at the top and four ER1.5
at the bottom; see Figure 21. Remove the cage rods’ setscrews that are pointing away
from the beamsplitter cube. Attach an SM1CP2 end cap to the left side of the CCM1-
4ER(/M) cube.
Beamsplitter Cube

Objective
Figure 20 shows the components of the objective assembly. First, remove the retaining
rings from the CP02(/M) cage plate and attach the objective to it using the SM1A17
adapter. Then, mount the cage plate to the C1498(/M) post clamp using an SD1
counterbore adapter and an 8-32, 1/2" long (M4 x 12 mm) cap screw. Make sure not to
mistake the thick SD1 adapter with a thin washer. Also, the C1498(/M)’s label should face
upwards. Ensure that you do not touch the bottom of the objective with your fingers, in
order to prevent possible contamination.
Microscope Objective
Assembled
Components
Microscope Objective
Slip the objective’s cage plate onto the cage rods below the beamsplitter, at about half
length of the cage rods. Tighten the plate’s locking screws to secure the objective. Make
sure to handle the objective with care and do not lay it on the table with the optic facing
downward.
Page 34 MTN012639-D02
LED Assembly
The components for the LED assembly are shown in Figure 22. First, remove the two
retaining rings from the CP02(/M) cage plate and attach the MCWHL5 LED to it using the
lens tube coupler (SM1T2). Connect the SM1L10 lens tube to the CP02(/M) cage plate.
Mount the cage plate to the C1498(/M) post clamp using an SD1 counterbore adapter and
an 8-32, 1/2" long (M4 x 12 mm) cap screw. Finally, install the DG10-600 diffuser. The
diffuser is supposed to sit at the very top of the SM1L10 lens tube. For that, the retaining
ring needs to be positioned close to the edge. Place the DG10-600 on top. Take one of
the two retaining rings from the CP02(/M) and fix the DG10-600 with it.
LED Components

Camera System
Figure 23 shows the components for the lens 3 assembly. Mount the LB1676 lens to the
CP02(/M) cage plate using two retaining rings.
Lens 3 Components
Next, attach the FES0650 bandpass filter to the CP02T(/M) thick cage plate. Figure 24
shows the necessary components. Note that the filter has a defined direction of light
propagation. This is marked on the frame of the filter with a small white arrow. This arrow
should be pointing up during assembly.
Bandpass Filter Components

Figure 25 shows the components for assembling the top part of the camera system.
 First, install the two FGB37 filters in the SM1L15 lens tube using retaining rings.
Screw one retaining ring into the lens tube about half-way, then insert the first
filter, and then secure it with a second retaining ring. Insert the second filter, and
then insert a third retaining ring (unused from one of the cage plates) to secure
it.
 Second, connect the DCC1645C camera to the SM1T2 lens tube coupler using
the included adapter (CS mount to internal SM1). After this, screw the other side
of the lens tube onto the SPT1(/M) slip plate positioner (with the locking screws
facing towards the camera). The SM1T2 coupler allows the user to choose the
camera orientation with the Thorlabs logo facing to the right, see Figure 26, so it
is parallel to the stage orientation.
Page 36 MTN012639-D02
 Third, connect the CP02(/M) cage plate to the C1498(/M) post clamp using an
SD1 counterbore adapter and an 8-32, 1/2" long (M4 x 12 mm) cap screw.
 Finally, place the SPT1(/M) and CP02(/M) onto each other and slip four ER3 cage
rods through them (with the rods’ setscrews removed on both sides). Ideally, the
cage rods ends should be flush with the upper side of the SPT1(/M). Attach them
both to the rods by tightening the small cage locking screws at the side.
Camera System Components

Slip lens 3 from Figure 23 onto the cage
rods and secure it with the locking
screws on the side. Make sure the
locking screws do not point towards the
side where the damped post will be (i.e.,
the side of the C1498(/M) clamp).
Next, slip on the filter in the cage plate
from Figure 24. Also make sure no
locking screws face towards the
C1498(/M) clamp. The cage rods should
be inserted into the CP02T(/M) only half
way. Make sure that the upper locking
screws fix the cage rods to the thick cage
plate. The lower locking screws need to
be left free (so the camera segment can
be attached to the beamsplitter
assembly). The fully assembly camera
segment can be seen in Figure 26.
Camera System

Microscope Assembly
 Attach the LED assembly to the damped post. The
LED can be in contact with the breadboard. Try to
align the cage plate of the LED assembly parallel
to the breadboard hole pattern.
 Attach the objective assembly to the damped post.
The distance from breadboard top to the bottom of
the objective should be about 11.5 cm.
Use a level at the beamsplitter cube to make
sure the assembly is vertical. If not, you have to
adjust the angle in which the C1498(/M) clamp is
screwed to the cage plate.
 Put the camera assembly on top of the objective
assembly. The short cage rods on top of the
beamsplitter should fit easily into the thick cage
plate at the bottom of the camera assembly. If not,
adjust the angle at which the C1498(/M) clamp is
screwed to the respective cage plate.
The microscope assembly is now complete and should look
like the image on the right.
Page 38 MTN012639-D02
Next, connect the laser assembly and the microscope. For that, screw the remaining
SM1L10 into the side of the beamsplitter. Then, adjust the position and height of the
KCB1(/M) right-angle kinematic mirror mount by loosening the SM1CLP10 flexure sleeve
couplers. When they are in place, make sure the KCB1(/M) are still parallel to each other
(to check that, you can use a flat object like a book or level). Clamp the posts supporting
the cage segment and the laser to the breadboard using the two CF125 clamps and two
1/4"-20, 3/8" long (M6 x 10 mm) cap screws with washers.
Note: If the objective/microscope was assembled with a height that differs strongly from
the aforementioned 11.5 cm (distance from the breadboard to bottom of the objective), the
SM1CPL10 might not be long enough to cover both tubes. In that case, you need to change
the height of the entire microscope by moving the objective assembly and the camera
assembly either up or down.
The optical tweezers are now complete except for the sample positioning table and the
control elements. Figure 27 shows the current assembly.
Complete Trap Laser and Microscope System

6.1.4. Sample Positioning System

Like the previous units, the control and positioning unit consists of several individual
subassemblies, which are constructed first.
First, take the MT401(/M) base plate and add
4 dowel pins.
In this setup, we use the MT405 to side-

mount the motors to the MT1(/M) stage. Do
the following for the two MT1(/M)-Z8 stages:
First, take the 3/32" hex key from the MT405.
Remove the end-mount adapter by removing
the two inner screws.
Do not remove the locking plate and attempt

to mount an actuator on that side of the stage.
First, screw the two parts of the MT405 to the
side of the stage. The hollow part that holds
the barrel/motor is mounted to the thin, lower
part of the stage. The part with the silver inset
is screwed to the thick, upper part of the
stage. Make sure to tighten the respective
screws symmetrically. Insert the motor and
tighten the clamping screw.
Customers with the metric kit should make
sure to label and store the MT405’s 3/32"
(imperial) hex key for future use.
Page 40 MTN012639-D02
Screw the first MT1(/M)-Z8 with side-

mounted motor to the base plate in the
orientation shown to the right. Use two 1/4"-
20, 3/8" long (M6 x 10 mm) screws.
Insert 2 dowel pins into the top of the stage.
Screw the second MT1(/M)-Z8

with side-mounted motor onto
the first one in the orientation
shown in the image to the right
using two 1/4"-20, 3/8" long
(M6 x 10 mm) cap screws.
Add 2 dowel pins to the top.
Screw the MT402 right angle

bracket to the stage assembly
using two 1/4"-20, 5/8" long
(M6 x 16 mm) cap screws with
washers.
Add 2 dowel pins to the thick
side of the MT1B(/M) stage and
screw it to the MT402 using two
1/4"-20, 5/8" long (M6 x 16 mm)
cap screws with washers.

Connect the two pieces of the

sample holder using two 8-32",
3/8" long (M4 x 10) mm screws
with washers.
Screw the adapter assembly to

the stage using two 1/4"-20,
0.315" long (M6 x 8 mm) cap
screws with washers and 4
dowel pins (you may need to
use the screws to pull the
stages together once you have
aligned the dowel pins).
Now, attach the assembled stage on the breadboard at the position depicted in Figure 10
using four 1/4"-20, 3/8" long (M6 x 10 mm) cap screws with washers.
Page 42 MTN012639-D02
The next step is to install the electronic controls, see Figure 28, onto the breadboard.
2 x KDC101, LEDD1B, and KLD101 Controllers

First, connect the LEDD1B LED Driver to the MCWHL5 LED and position it on the
breadboard using two 1/4"-20, 1/2" long (M6 x 12 mm) cap screws with washers (the
same type of screws and washers should be used with the following controllers as well).
Then, place the two KDC101 K-Cube Stage controller cubes in their positions, mount
them to the breadboard, and connect them to the computer using USB cables. Next,
connect the KLD101 laser diode controller to the SR9A-DB9 mount, and mount it on the
breadboard. In addition, you must install the supplied software programs for the
operation of the DCC1645C camera and the cubes (KDC101, KLD101). Further
adjustment of the camera and beam path are required for operation. You can find the
instructions for this in Sections 6.3 to 6.5.

6.2. Controller and Software Setup

6.2.1. LED Controller
The front of the LED Controller has a current limit adjuster. Set the
maximum current of 1 A when using the MCWHL5 LED. The mode
switch on top of the controller should be set to CW.
6.2.2. Laser Controller

Install the Kinesis control software, which will allow for control of the laser diode controller
and sample positioning stages. The software can be installed using the provided CD or
downloaded from the Thorlabs website. Connect the laser controller to the PC only after
you are asked to during the installation. Afterwards, open the newly installed program.
Kinesis Laser Controller Panel.

In the Kinesis control panel, see Figure 29, press “Settings”.
 To operate the laser properly, the polarity setting of the diode will need to be
switched to anode grounded in the “Control” tab. For that, select “Anode” in the
drop-down menu labeled “Polarity”, see Figure 30.
 Set the maximum current for safe operation of the diode in the tab “Max Current”
to 100 mA, see Figure 30. By activating the “Persist Settings to Hardware”
checkbox, you can store the settings on the device.
Page 44 MTN012639-D02
 It is possible to monitor the laser diode with the included photodiode, see Figure
31. This is not necessary for this kit and should not be enabled.
 Once the polarity and the maximum current are set and persisted to the hardware,
you can operate the laser driver with the controls on the controller itself (rather
than controlling it by software). The current can be set by turning the
potentiometer knob at the top of the controller. For that, the “Advanced” settings
should look like in Figure 31.
The current range of the L658P040 laser diode is 75 mA (typical) to 110 mA
(maximum). The actual output for a certain current depends on the diode at hand.
You have two options:
 Set the current to approximately 80 mA and measure the output power with
a power meter. Adjust the voltage until you reach 40 mW of output power.
 Set the operating current just below 100 mA. According to our test
measurements, this will result in an output power above 50 mW. The
advantage is a stronger optical trap, the downside is that operating a laser
diode above the specified output power may decrease its lifetime.8
Settings of the Laser Diode Controller
8
So far, we have not seen a significantly increased failure rate with an operating current near
100 mA. However, we cannot guarantee that the diode reaches the typical lifetime when driven near
its current limit.
More Settings for the Laser Diode Controller

Figure 32 shows the Kinesis software with the laser switched on and a current of 95.0 mA.
Kinesis Software for Laser Diode Controller with Laser Switched On
Page 46 MTN012639-D02
6.2.3. Camera
To operate the camera download the latest ThorCam Software from the Thorlabs website
or use the installation CD included with the camera.9 Follow the installation guide and
select DCU/DCC Series Compact CMOS and CCD Cameras. After finishing the installation
reboot and connect the camera to your computer. After the driver initialization you can start
the ThorCam Software.
ThorCam Start Panel

Choose the camera you want to activate and press the start button (see Figure 33). This
will start the software front panel with a black screen shown in Figure 34
Home Screen of the ThorCam DCC Front Panel
9
https://www.thorlabs.com/software_pages/ViewSoftwarePage.cfm?Code=ThorCam
To see live transmission from the camera, select Start Capture from the command
panel. You can change the brightness of the LED to check the signal coming from the
camera.
A helpful feature is the “Draw Circle” option .You can use it to mark the location of
your laser spot. This can be helpful when you move out of the trapping plane but still want
to know the location of the laser focus. An example can be seen in Figure 35. There is also
the possibility to label the laser focus using the text insert function via the button .
“Draw Circle” Option to Mark the Location of the Laser Spot

The image properties need to be changed so that we can observe the laser spot later on.
To do so, open the Settings to open the window shown in Figure 36.
Settings of ThorCam DCC Camera Software
Page 48 MTN012639-D02
In the Camera tab you can choose the Pixel Clock, Frame Rate, and Exposure Time. The
basic parameter for camera timing is the pixel clock. It determines the speed at which the
sensor pixels are read out.
We recommend setting the pixel clock no higher than necessary to achieve the desired
frame rate. Adjust the pixel clock while observing the percentage of dropped frames shown
beneath the Pixel Clock slider.
Frame Rate: The possible range of settings for the frame rate depends on the currently
selected pixel clock. You can select a lower frame rate without changing the pixel clock.
To set a higher frame rate, however, the pixel clock might need to be increased. For the
optical tweezers application, we recommend a frame rate of 10 FPS or higher.
Exposure Time: The exposure time depends on the currently selected frame rate and is
preset to its reciprocal value. A shorter exposure time can be set without changing the
frame rate. A longer exposure time will reduce the frame rate.
For image improvements adjust the settings in the Image tab. The recommended settings
are shown in Figure 37. The high red gain compensates for the red bandpass filter in front
of the camera (to avoid laser damage). You may reduce the red gain when you have found
a pronounced focus. Also, you can simply use the Auto White Balance function (and
increase the red gain afterwards).
Image Settings
With the ThorCam software you can also save snapshots during live imaging or
record videos. The recording dialog box shown in Figure 38 gives you the option
of limiting the recording size by setting the maximum number of frames or the maximum
file size in MB.
Record Video Settings

For further information on the settings check the ThorCam user guide installed in the
program’s folder.
6.2.4. Sample Positioning Stages

The stage is driven by Z812B brushed motors, which are controlled by two KDC101
devices. For optical trapping, low driving speeds are necessary. To adjust the velocity of
the servo motors you can either use the display of the KDC101 or the configuration
software. You can find the software via the Thorlabs product page10 of either the KDC101
or the EDU-OT2(/M).
Method 1: Configuration directly via KDC101 display and wheel: Power up the
controller and open the menu on the KDC101 controller via the menu button, select “set
velocity” with the wheel and confirm with the menu button. You can now set the velocity to
the recommended 0.005 to 0.03 mm/s. Furthermore select “Joystick Mode” in the menu
and chose “Velocity Control”. This corresponds to a continuous motion of the stage when
the wheel is actuated. Confirm with the menu button.
Method 2: Configuration via Software: The controllers can also be configured via the
Kinesis software as shown for the laser diode controller. If not installed yet, download
Kinesis and follow the installation instructions. Connect the controllers via USB when
asked or after the complete installation.
Start the Kinesis software. The controllers are shown automatically. Check the device
serial numbers in the software and on the K-cubes.
10 www.thorlabs.com/newgrouppage9.cfm?objectgroup_ID=2419
Page 50 MTN012639-D02
The two windows, shown in Figure 39, correspond to the two servo motors. To establish
which window controls which controller cube, press the “Ident” or “Identify” button in the
software, which will flash the display a few times on the corresponding controller.
Kinesis Software User Interface

Settings:
 Press the Settings button in the Kinesis software and select the “Advanced” tab
(Figure 40).
 In the Device Controls field, choose “Velocity control” for the Wheel Mode
(sometimes also referred to as Joystick Mode). This corresponds to a continuous
motion of the stage when the wheel is actuated.
 The recommended Max. Velocity is set to 0.03 mm/s and Acceleration to 1 - 2
mm/s2.
 The recommended11 Drive Array Velocities are set to
o Velocity 4: 0.03
o Velocity 3: 0.02
o Velocity 2: 0.01
o Velocity 1: 0.005
This defines the different velocities which can be accessed via drive buttons in
the software as shown in Figure 41. The controller wheel deflection is proportional
to the max. velocity.
 The settings need to be entered for each servo motor separately. All other
settings remain default values.
Important Note: By clicking OK, the settings are sent to the device. By checking the
Persist Settings to the Device checkbox the settings are stored in the device and
11
These velocities are good start values to ensure that the particles are not lost when moving at
maximum speed. If the Optical Tweezer is adjusted very well, trapping is possible at even higher
velocities.
loaded after each restart of the controllers. In the Kinesis Software you can also enter
all parameters under the Device Startup Settings tab. By clicking Save, this will store
the settings on your computer as well. When the controllers are reconnected to this
computer they will download the stored settings immediately. You can find further
information in the KDC101 manual.
You can operate the motors either by hand via the controllers or by software. When you
choose the latter, you can move the stage by pressing the “Drive” buttons in the main
window, see Figure 41. Here, you can also open the Drive Array Velocities in a drop-down
menu. Each of the horizontal blue bars in the “Drive” button field corresponds to one of the
velocities.
Kinesis KDC101 Settings
Kinesis Drive Array Velocity

Page 52 MTN012639-D02
6.2.5. Kinesis Settings

Thorlabs Kinesis software can be set to grab the current settings from the connected
devices on startup. This option can be activated via the dropdown menu File -> Options
by checking the box Use device persisted settings (see Figure 42).
Use Device Persisted Settings

In most cases for the optical tweezers setup you would want to read the settings of the
device, so before connecting the controllers, open the Kinesis software and select Use
device persisted settings. This will be applied for all connected devices.

6.3. Camera Adjustment

The objective used in this kit is infinity-corrected. That is a great advantage and eases the
adjustment significantly. It means that the light from the objective’s focal plane leaves the
objective effectively parallel. This plane can be mapped onto a camera with a lens at an
almost arbitrary distance to the objective. To find the correct distance between lens and
camera chip, one only needs to map a distant object onto the chip. Proceed the following
way:
Step 1: Preparation
 Remove the camera segment from the microscope (that includes all parts starting
from the camera until the cage plate above the beamsplitter). For that, unscrew
the cage locking screws from the CP02T(/M) and loosen the screw on the
C1498(/M) post clamp.
 Start the camera software.
 Open the settings and set the gain to maximum.
Step 2: Lens Adjustment

 Aim the camera at an object more than
4 m away. Make sure the room is well
lit or activate the gain boost in the
camera settings (beneath the regular
gain). You could also increase the
exposure time.
 Move the lens in the cage segment until
you see a sharp image of the object.
Below is a sample image of a clock at 4
meters distance.
Step 3: Installation of the camera

 Place the camera segment back into the microscope. Tighten the cage locking
screws of the CP02T(/M) and fix the screw on the C1498(/M) post clamp.
Page 54 MTN012639-D02
6.4. Beam Adjustment
Warning
The class 3B laser diode used in this kit can emit more than 50 mW of optical power,
which can cause damage to the eyes if viewed directly. The laser driver is equipped with
a key switch and safety interlock, which should be used appropriately to avoid injury.
Additionally, we recommend wearing appropriate laser safety glasses
when using this kit.
Step 1: Preparation
 Remove the removable cage segment with the two lenses forming the beam
expander.
 Take the laser out of its
mount by opening the
locking screws on the
holder. Guide it through the
CP09(/M) cage plate.
 Screw in the collimation lens’
retaining ring as far as
possible.
 Screw in the collimation lens
as far as possible (as close
to the diode as possible),
see image to the right.
Step 2: Collimation
 Turn the safety key on the KLD101 laser driver to ON.
 Press the “Laser On” button.
 Turn the knob to set the diode current to 50 mA.12
 After ensuring that the area is clear of other people, point the laser at a wall,
preferably more than 3 m away.
 Slowly screw the collimation lens away from the diode until you see the laser light
distribution on the wall has turned into a small spot. Note that the pattern will be
slightly asymmetrical. That is not a problem.
 Fix the collimation lens with the collimator’s locking ring.
 Turn off the laser by pressing the button near the “Laser On” light once again.
12
Make sure you have set the maximum current to protect the laser diode, see Chapter 6.2.2.
Step 3: Laser Installation

 Place the alignment plate into the cage system
right before the 45° mirror, see image to the right.
 Insert the laser into the mount without securing it
with the locking screws yet.
 Turn the laser on and observe the pattern on the
alignment target. You will see an elliptical shape.
 Rotate the collimator in the housing until the long
side of the ellipse is oriented in vertical direction.
 Fix the locking screws. Make sure the locking
screws fix the collimator close to the thread, see
image below.
Step 4: Laser Mount Alignment

 Turn the tip and tilt screws on the KC1-T mount
until the brightest spot of the laser light
distribution hits the hole in the alignment plate.
 Turn off the laser.
 Remove the alignment plate.
Step 5: Adjustment Mirror Alignment, Part 1

 The second (upper) adjustment mirror
must also be removed from its mount.
To do this unscrew the set screw,
place a piece of adhesive tape on the
back of the mirror and pull it out of the
mount.
 Turn on the laser.
 Adjust the lower adjustment mirror so
the laser beam exits the upper KCB1
vertically. For that, using a piece of
paper to view the laser spot helps.
When aligned vertically, the laser beam will exit close to one side of the mount
aperture. That is not problematic: when the mirror is installed, the laser will hit the
mirror approximately in the center.
 Place the mirror back into the mount and fix the locking screw.
Page 56 MTN012639-D02
Step 6: Adjustment Mirror Alignment, Part 2

 Remove the end cap on the beamsplitter.
 Turn on the laser.
 Adjust the upper alignment mirror so that the
laser exits the beamsplitter at its center.
 Screw the end cap back on.
Step 7: Insert Beam Expander

 Adjust the distance between the lenses: the
distance between the CP09(/M) cage plates
should be 12 cm (distance between adjacent edges).
 Place the beam expander section back into the cage segment.
 Turn on the laser. It should look like the photo below. Note that the elliptical beam
is oriented with the long arm vertical.

Step 8: Final Adjustment

 Increase the diode current to approximately
95 mA.
 Observe the expanded laser spot on the
objective aperture. Adjust the upper alignment
mirror to move the laser spot into the center of
the objective aperture.
Note: Ideally, an optical tweezers system consists of a single mode laser that enters the
objective as centered and vertical as possible. Also, the laser beam should be as parallel
as possible, with a small divergence angle. In our setup, this is not the case: After following
the instructions above, remove the end cap at the beamsplitter and examine the beam
shape using a viewing card or piece of paper. Notice that it converges at a distance of
about 1 to 2 meters. In other words, the light is not very parallel.
However, keep in mind that we do not have a single mode laser. Since we optimized the
setup for low cost, the laser diode used is relatively cheap. The intensity distribution of the
laser diode is elliptical and differs quite a bit from the ideal, Gaussian case. In our trials,
the trapping was clearly better when the laser showed the described convergence.
Therefore, the adjustment instructions above were written to yield optimal trapping results
rather than a minimum divergence angle.
Page 58 MTN012639-D02
6.5. Trapping, Microscope Focus, and Final Alignment

The table is now set to trap microscopic particles.
Step 1: Preparation
 Switch on your computer and start the camera software (see Section 6.2.3).
 Set the velocity of the motors to 0.02 mm/s (see Section 6.2.4, you may increase
the speed when you have trapped a particle).
 Set the LED lighting to maximum (see Section 6.2.1).
 Switch on the laser with a diode current of 95 mA (or the diode current you
measured to obtain 40 mW of output power, see Section 6.2.2)
Step 2: Sample
 Prepare a sample with polystyrene-beads (see Section 7.1).
 Place the sample positioning table as low as necessary with the aid of the
micrometer screw for the Z direction. Make sure to leave enough space to avoid
a collision with the objective when the sample slide is inserted into its holder.
 Place the sample in the holder with the selected well under the objective.
Step 3: Approach
 Raise the sample positioning table with the micrometer screw and pay close
attention to the camera image on the monitor of your computer. Note that the
objective’s working distance is below 0.5 mm due to the high numerical aperture.
 As you raise the sample, you will see a distorted red laser spot forming on the
monitor. This is not the sample, but rather the top of the cover glass. Continue
slowly raising the sample. The laser spot will disappear and reappear. The
second time you see the spot indicates the bottom of the cover glass. Raise the
sample higher, and the spot will disappear and reappear again. The third time
you see the spot indicates that the laser focus is in the sample. At this point, the
focus is at the right level. Figure 43 shows a schematic representation of the
situations.
Please note that reflections of the laser may occur. In the camera image, these will appear
with varying diameter (small spots or larger shapes). These are not a sign of bad
adjustment and can be ignored.

Focus Level Settings

 The camera image should look similar to Figure 44.
Camera Image when the Sample is Approached (2 µm Beads)
Step 4: Camera Position

 Depending on the angle of the optical elements, the laser spot may not be in the
center of the camera image. In fact, this is hardly ever the case and does NOT
constitute bad alignment.
 You can move the laser spot closer to the center of the camera image by
physically moving the camera. The camera’s lens tube is mounted on an SPT1
slip plate that allows travel of ± 1 mm in each direction. Loosen the two screws at
the top, move the slip plate with the attached lens tube/camera and fix the screws
again.
 In rare cases, you may initially not see a laser spot in the camera image at all. In
these cases, you may find it within the travel range of the SPT1. Make sure you
have the correct plane by getting the beads in focus and move the SPT1’s slip
plate to find the laser spot.
Page 60 MTN012639-D02
Step 5: Trap a Bead

 Move the stage using the wheels on the controller cubes. Move the sample so
that a bead comes to rest under the laser spot, see Figure 45.
 When the bead is close to the spot, you should see it getting pulled into the focus.
 Move the sample around by moving the stage. The bead should stay trapped in
the laser focus while the sample is moved.
Bead is Trapped in the Laser Focus
Step 6: Final Corrections and Potential Errors

 If you still have trouble seeing the laser spot, set the red amplification to maximum
when adjusting the live image. You can also try dimming the LED.
 If trapping is weak, slightly move the sample up or down to find the optimal plane
of trapping.
 If the laser spot is in a significantly different plane than the plane where the beads
can be seen sharply, you may have to adjust the focal planes, see Figure 46 and
the explanation below. Move the lens in front of the camera in the Z direction
along the cage rods until you have a sharp image of both the beads and the laser
spot. If beads are pulled out of the focal plane when trapped, the focal planes do
not coincide.
 If you want the movement in Z direction to be smoother, a hex key can be inserted
in the adjuster of the MT1B. This allows you to move the stage up and down with
less vibration in the system.

Laser Focal Plane vs. Microscope Focal Plane

The position of the laser focus in the Z direction is defined by the focal length of the
objective (red curve in Figure 46). For ideal observation of both the laser focus and the
trapped sample beads it is necessary to adjust the microscope (tube) lens (shown in Figure
46) in such a way that the focal plane of the observing camera system (green curve) -
consisting of the objective, the tube lens and the CCD camera chip - coincides with the
fixed focal plane of the laser. When adjusted correctly, the focused laser spot, as well as
the sample beads in the plane of the focus, are both visible through the microscope.
Camera
Microscope Lens
Beamsplitter Laser
Objective
Focal Plane Camera

Focal Plane Laser
Laser and Camera Focal Planes
Page 62 MTN012639-D02
Portable Optical Tweezers Kit Chapter 7: Experiments
Chapter 7 Experiments
In this chapter, we discuss exercises using glass beads as samples. The material can be
found in the OTKBTK preparation kit, which is sold separately on the Thorlabs webpage.
However, these experiments can also be conducted with several other materials, including
polystyrene beads or lipids in aqueous solution.
We do, however, recommend the use of the OTKBTK since the setup was optimized for
the sample slides and the cover glasses provided with the OTKBTK.
Content of the OTKBTK Preparation Kit with tools for glass bead
sample preparation (green highlighting). Do not use immersion oil.
7.1. Creating a Sample

A sample containing 1 µm or 3 µm silica beads is useful, as these are well-suited for getting
to know the operation and handling of the optical tweezers. The following materials are
necessary to create the sample:
 Microscope slide with around 20 µm deep cutouts
 Cover glass
 Pipette
 Solution with fused silica (glass) beads
 Do NOT use immersion oil. The supplied objective is an air objective and
immersion oil is neither needed nor recommended in this setup.

First, shake the glass bead solution and place one or two droplets (approx. 30-40 µl) in a
cutout on the microscope slide using a pipette. Place a cover glass over the sample,
starting with the glass tilted at a steep angle and lowering it so that there are no air bubbles
between the glass and the sample (see Figure 48). Remove excess solution with a tissue.
Droplet of Bead Solution on Glass Slide

The samples can either be prepared before each experiment or they can be sealed
between the slide and the coverglass with a UV adhesive. We recommend allowing
students to prepare new samples as an educational exercise.13
7.2. Setting the Correct Focus Level

Place the sample positioning table as low as necessary using the micrometer screw on the
z-axis stage and place the selected sample under the objective. Move the sample stage
to center the sample under the objective. For this step, it may be convenient to temporarily
set the maximum velocity of the KDC101 controllers to a higher value, see Section 6.2.4.
Now, raise the sample positioning table once again using the micrometer screw and pay
close attention to the camera image on the monitor of your computer.
After a certain distance, you will see a laser spot in the form of a red, distorted spot on the
monitor. This is not the sample, but rather the top of the cover glass. The second visible
spot, seen after moving the objective closer to the sample, shows the bottom of the cover
glass. Moving the objective closer to the sample, the third visible spot is inside the sample.
Once this is in view, the focus is at the right level. Figure 49 shows a schematic
representation of the laser focus levels.
Note: It is possible that large red laser reflections are observed which do not change even
when the z-axis (focal plane) is changed. They originate from the beamsplitter and have
nothing to do with the laser spot of the optical trap. Also, do not move the sample
positioning table too far upwards. Otherwise, there is a risk of destroying the cover glass
by touching it with the objective. This will damage the objective and would entail
unnecessary cleaning work.
13
Replacement pipette tips can be found here http://www.accumaximum.com/tips-ordering.html
Page 64 MTN012639-D02
Focus Level Settings

Set the laser focus so that you see the sample beads clearly and sharply. By moving the
laser spot onto a bead using the servomotors and levers on the K-cube controllers, you
can trap a bead and move it to another location. If you are able to capture one of the beads,
you have found the correct focus level.
Please note that reflections of the laser may occur. In the camera image, these will appear
with varying diameter (small spots or larger shapes). These are not a sign of bad
adjustment and can be ignored.
At this point, if the trapped beads move out of the focal plane, it indicates that the focal
planes of the laser and camera system are not aligned. See Section 6.5, above, for details.
7.3. Arranging the Silica Beads

In order to get to know the optical tweezers better, try next to move as many beads as
possible to one location using the K-cube controllers as shown in Figure 50. These
controllers allow you to set different speeds. You can use this to your advantage when
releasing the beads.
If you experience lagging of the camera image, try to reduce the frame rate in the camera
settings.
Hint
With older or slightly dried samples, one can observe that some beads no longer move,
but instead "stick" in place.

Beads as Viewed Through the Microscope
7.4. Manipulating a Dairy Cream Particle in a Cream/Water

Emulsion
Mix a drop of dairy cream with sufficient water so that the solution is still slightly milky.
Now, create a finished sample for the optical tweezers as described in Section 7.1, above.
Observe the solution under the optical tweezers.
Exercise
What do you observe? Try to bring a particle into the beam path. What happens? Then,
try to track the particle by changing the z- position. Where is it? Switch the laser off and
observe the particle. What is happening here?
Solution
In this part of the experiment, a sample must first be created out of a cream/water
emulsion. If one attempts to trap the cream particles with the laser, it is observed that they
disappear from the focus and can no longer be clearly seen on the monitor (see Figure
51). The explanation for this observation can be found by examining more closely the
cream/water emulsion. Cream consists primarily of fat, which collects on the surface when
mixed with water. The cream particles, which are trapped by the laser, are therefore
located on the surface of the water. However, the laser focus is located at a deeper level,
so when the cream particles are trapped, they are pulled down into the emulsion. This
effect can be observed when the particle at which the laser is directed is tracked with the
aid of the z-axis screw.
Page 66 MTN012639-D02
Cream Particles in a Cream / Water Emulsion

If a cream particle, which is located in the optical trap, has been brought into focus with
the z-axis screw and thus can clearly be seen on the monitor, the laser can be switched
off and the particle observed. Switching off the laser causes the cream particle to no longer
be located in the optical trap and thus to move upward once again to the surface of the
water. This procedure can be observed again with the aid of the z-axis screw.
Cream particles collect on the surface of the

water.

A cream particle is pushed down by the laser

focus.
Cream particle is trapped at the laser’s

focus.
Once the laser is switched off, the fat particle

moves back to the surface of the water.
Page 68 MTN012639-D02
7.5. The Holding Force of the Optical Trap
7.5.1. Brownian Motion

The proper motion or vibrating motion of particles, also referred to as Brownian motion,
can be observed under the optical tweezers. Samples can be found at
www.polysciences.com “Microspheres and Particles”.
Exercise
Explain briefly the causes of Brownian motion and what role it plays in the experiments
with the optical tweezers.
Solution
Brownian motion is the statistic vibrating motion (translation and rotation) of microscopic
free particles. Under the microscope, the paths of particles are seen as short, straight lines
(see Figure 52). The Brownian motion can be observed in experiments under the optical
tweezers. The polystyrene beads are located in a medium that consist of molecules. These
molecules are constantly moving in all directions. Because of this, the molecules
repeatedly bump into the beads, which causes a vibrating motion of the beads that can be
observed under the optical tweezers. The molecules move more the higher the
temperature.
Brownian Motion
Use the sample with the 3 µm polystyrene spheres. You must first switch the laser off so
that you can observe only Brownian motion. For evaluation, a video sequence with a
duration of 2 minutes or more must be recorded. During this period, about 5 particles,
which do not touch each other, should be in the image. A similar video should be recorded
with the 1 µm spheres. The videos must be evaluated with the aid of image analysis
software, which provides the x and y position of a particle over time. A program that makes
evaluation possible and is offered for free is “Viana”, available at
http://www.viananet.de/en.
Note: Before taking the video sequence, make sure that the beads do not show a drift in
any direction, which can be caused by an air bubble. If necessary, prepare another sample.
We recommend evaluating the data obtained with the aid of a table calculation program
and to show the results in a graph. First, the mean squared displacement 𝑟 2 (𝑡𝑖 ) of the
beads must be determined. This can be calculated with the positions (𝑥𝑖 (𝑡𝑖 ), 𝑦𝑖 (𝑡𝑖 )) of the
particles at different times 𝑡𝑖 , which must be taken from the video:
2 2
𝑟 2 (𝑡𝑖 ) = (𝑥(𝑡𝑖 ) − 𝑥(0)) + (𝑦(𝑡𝑖 ) − 𝑦(0)) (29)
The time mean value 〈𝑟 2 〉𝑡𝑖𝑚𝑒 at time 𝑡𝑛 results from the averaging of all measured values
𝑟 2 (𝑡1 ), … , 𝑟 2 (𝑡𝑛 ) at the measurement times:
𝑛
1
〈𝑟 2 〉 2
𝑡𝑖𝑚𝑒 (𝑡𝑛 ) = ∑ 𝑟 (𝑡𝑖 ) (30)
𝑛
𝑖=1
In order to eliminate statistically possible deviations of individual particles, it is

recommended to once again determine the mean value over 𝑀 various particles. We
recommend doing this for at least 5 particles.
𝑛
1 1
〈𝑟 2 〉(𝑡𝑛 ) = ∑ ∑ 𝑟 2 (𝑡𝑖 ) (31)
𝑀 𝑛
𝑖=1
The obtained values for average displacement, 〈𝑟 2 〉(𝑡𝑛 ), are now plotted with respect to
time. Figure 53 shows an example of mean squared displacement over time for three
different sizes of polystyrene beads. Here, each straight line is the mean value of several
particles of the same sample. It is recommended to perform a linear fit through the resulting
curves in order to obtain the slope, 𝑚, of the straight lines.
Exercise
What differences do you expect between the Brownian motion of the 3 µm and 1 µm
spheres and why?
Solution
The slope of the lines in Figure 53 decreases with increasing diameter of the beads,
meaning that larger beads move less. This result can be easily explained through
Brownian motion. The 1 µm spheres can be more easily sent into motion by impact with
the water molecules than larger spheres. Therefore, a 1 µm bead travels more in a certain
time interval than a larger bead.
Page 70 MTN012639-D02
Mean Squared Displacement for Polystyrene Beads of Different

Sizes
7.5.2. The Maximum Holding Force

In the following, we shall determine how well the optical tweezers can hold a polystyrene
bead. The goal is to determine the maximum holding force of the optical trap.
If one observes an individual polystyrene bead under the microscope which is moving
freely, frictional force acts on the particle. This inhibits the particle in its movement through
any liquid. This frictional force 𝐹𝑅 is directly proportional to the speed 𝑣, with which the
spheres move and can be described by the following equation:
𝐹𝑅 = 6𝜋𝜂𝑒𝑓𝑓 𝑅𝑣 (32)
Here, 𝑅 describes the radius of the bead and 𝜂𝑒𝑓𝑓 describes the effect of viscosity of the
suspension. The latter indicates how "thick" the combination of water and beads is and is
different for each sample. As the viscosity depends on various factors, it must be
determined through experimentation. It depends upon the mean squared displacement of
the particles, which was determined in the previous exercise. This resulted in a straight
line, whose gradient can be determined by means of the following equation:
2𝑘𝐵 𝑇
𝑚= (33)
3𝜋𝜂𝑒𝑓𝑓 𝑅

Here, 𝜂𝑒𝑓𝑓 denotes the effective viscosity, 𝑅 is the radius of the PS-bead, 𝑇 is the
temperature of the sample in Kelvin (corresponds to room temperature), and 𝑘𝐵 is the
𝐽
Boltzmann constant, which is a natural constant and has a value of 1.38 ⋅ 10−23 𝐾 .
Exercise
Determine the effective viscosity 𝜂𝑒𝑓𝑓 of the sample with the 3 µm polystyrene spheres, by
solving the equation (33) according to viscosity and using the gradient 𝑚 of the measured
curve from the previous exercise.
Solution
The equation for the calculation of the effective viscosity 𝜂𝑒𝑓𝑓 is:
2𝑘𝐵 𝑇
𝜂𝑒𝑓𝑓 = (34)
3𝜋𝑅𝑚
Here, T is the room temperature, kB the Boltzmann constant, a the radius, and m the
previously determined gradient of the PS-beads used.
𝑁𝑠
The determined effective viscosity should be in the range of a few 10−3 𝑚2
Exercise
After which speed can the PS-bead no longer be held? Determine the maximum holding
force of the optical tweezers.
Solution
If the PS-bead is in the optical trap, two forces act on it. First of all, the frictional force 𝐹𝑅 ,
which is caused by the suspension in which the PS-bead is located, and thus works against
the other force, the holding force 𝐹𝐻 of the optical trap. The following equation describes
the frictional force 𝐹𝑅 :
𝐹𝑅 = 6𝜋𝜂𝑒𝑓𝑓 𝑅𝑣 (35)
Here, 𝜂𝑒𝑓𝑓 is the effective viscosity of the suspension, 𝑅 is the radius of the bead, and 𝑣 is
the speed. The maximum holding force is said to have been reached precisely, when the
PS-bead at a certain speed 𝑣𝑚𝑎𝑥 can just be held. This is the case when both forces are
in balance:
𝐹𝑆𝑡𝑜𝑘𝑒𝑠 = 𝐹𝐻,𝑚𝑎𝑥 = 6𝜋𝜂𝑒𝑓𝑓 𝑅𝑣𝑚𝑎𝑥 (36)
The holding force is in the range of a few pN.
Page 72 MTN012639-D02
Portable Optical Tweezers Kit Chapter 8: Teaching Tips
Chapter 8 Teaching Tips

In this chapter we present a simplified approach to help students in understanding the
basic physical principles of optical tweezers. In the following, we discuss how a particle is
trapped in a focused laser beam and how a three-dimensional optical trap works.
For keeping an object in place, a restoring force has to act as soon as the object leaves
its equilibrium position. Since the object is trapped only by the laser, the restoring force
has to be applied by the laser itself.
An essential precondition for trapping an object is that it is transparent to the laser light.
This means that at least part of the light has to be transmitted. Part of the laser light is
reflected off the object’s surface, while the other part is refracted and transmitted. Thereby,
the laser faces a change in direction which corresponds to a change in the momentum.
This in turn means that there has to be a force between the laser light and the object.
Intuitively, one would expect the particle to be deflected. However, in an optical trap the
object is drawn into the laser focus.
The laser beam exhibits a certain intensity profile. In order to create an optical trap, this
profile needs to possess a point with maximal intensity, such as in a Gaussian profile.
Gaussian Intensity Profile

As shown in Figure 54, the cross section of a laser beam has a radius 𝑅. The intensity
varies radially: the intensity is maximal in the center of the beam and declines towards the
edge. Comparing a ray in the middle of the profile to a ray outside, the inner one shows
the maximal intensity while the outer one exhibits a lower intensity. This is demonstrated
by the thickness of the blue lines in Figure 54.
The entire laser beam is the sum of all rays. For simplicity, we will only consider two rays
in the following which are symmetrical around the beam axis. The insights we gain with
these two rays can be transferred to the entire beam.
First, we have a look at an unfocused (collimated) laser beam that is incident on a
polystyrene bead. We have to distinguish between two cases:
 The center of the bead is on the beam’s main axis
 The center of the bead is not on the beam’s main axis
Note: The size of the vector arrows in the following diagrams do not match from diagram
to diagram, and instead are enlarged where possible for clarity.
Unfocused Laser Beam, Particle on the Beam’s Main Axis:
We consider two partial beams which are symmetrically arranged around the beam axis,
as shown in Figure 55. The beams are labeled 𝑠 and 𝑡.
Unfocused Laser Beam Strikes a Polystyrene Bead Located on

the Beam Axis
Both partial beams have the same direction and intensity (since they both have the same
radial distance to the beam center with highest intensity) and carry the same momentum.
In Figure 56, the momentum vectors of the beams are depicted at some positions using
Page 74 MTN012639-D02
arrows in blue and green. The direction of the vector arrow corresponds to the direction of
the momentum and its length corresponds to the momentum’s magnitude.
Unfocused Laser Beam Momentum

For example, the vector labeled with 𝑝(𝑠, 1) depicts the momentum of the beam 𝑠 at
position 1. It important to note that the magnitude of the momentum, i.e., the length of the
arrows, does not change. However, the direction changes. In Figure 57, the vectors before
the entry of the beam into the sphere (position 1) and after the exit (position 3) are
subtracted to see how the momentum changes. The momentum change is depicted as
orange arrow.
Unfocused Laser Beam Momentum Changes

To obtain the net momentum change of both partial beams, we add both orange change
of momentum vectors in Figure 58. This line of argument can be transferred qualitatively
to the entire bundle of partial beams.
Unfocused Laser Beam Total Momentum Change

In Figure 58 we observe that the total laser beam experiences a momentum change in the
opposite direction of its direction of incidence. Momentum conservation then demands a
momentum change of the bead in downward direction. This means that the bead is pushed
away by the beam, along the optical axis. The beam experiences no momentum change
orthogonal to the optical axis, so it stays in the center of the beam.
Unfocused Laser Beam, Particle is not on the Beam’s Main Axis:
In the following, we follow the same train of thought as above but with the particle outside
the center of the beam. This is depicted in Figure 59.
Unfocused Laser Beam Incident on a Polystyrene Beam Located

Off the Beam Axis
Page 76 MTN012639-D02
In this case, the two partial beams do not feature the same intensity. Therefore, they do
not have the same momentum, as shown in Figure 60.
Unfocused Off-Axis Laser Beam Momentum

Both partial beams then exhibit the momentum change displayed in Figure 61.
Unfocused Off-Axis Laser Beam Momentum Changes

As shown, the partial beams exhibit momentum changes that differ in magnitude. Figure
62 shows the resulting net momentum change of the entire laser beam.

Unfocused Off-Axis Laser Beam Total Change in Momentum

Figure 62 demonstrates that the laser beam experiences a momentum change to the left
and upwards. This results in a momentum change of the bead to the right and downwards.
Therefore, the bead is drawn to the position with maximum intensity, forming a two-
dimensional trap. The bead, however, is still pushed downwards which means that it is not
trapped in once place.
Focused Laser Beam, Particle on the Beam’s Main Axis Below the Laser Focus:
We will next consider a focused laser beam. The particle is assumed to be in the center of
the beam but below the focus of the laser. The laser is assumed to be highly focused which
implies that the incident partial beams are no longer parallel.
Figure 63 depicts two partial beams of the focused laser. The center of the bead is
assumed to be below the focus.
Focused Laser Beam Incident on a Polystyrene Bead
Page 78 MTN012639-D02
Since the bead is in the geometrical center both partial beams have the same intensity
and, therefore, the same momentum amplitude. The direction off the momentum vectors
is different since the incident laser beam is focused.
Focused Laser Beam Momentum

Figure 65 displays the momentum change of both partial beams.
Focused Laser Beam Momentum Changes

The net momentum change of the entire beam is depicted in Figure 66.
Focused Laser Beam Change in Momentum

The laser beam exhibits a momentum change downwards. Hence, the bead’s momentum
is changed upwards. The bead moves against the beam’s main propagation direction and
into the laser focus. Analogously, one can show that the bead is pushed downwards when
the laser focus is beneath it.
In essence, a single laser beam can create an ideal three-dimensional optical trap. One
thing is crucial for functioning optical tweezers: in order to trap an object in a three-
dimensional trap, a highly focused laser beam with a suitable intensity profile is required.
The object is always pushed in direction towards the point with highest intensity:
 Radially into the middle of the laser beam due to the (Gaussian) intensity profile
 Vertically into the direction of the focus
As soon as a particle moves away from the focus, a restoring force acts on it to push it
back into the focus.
Page 80 MTN012639-D02
Portable Optical Tweezers Kit Chapter 9: Control via Game Controller
Chapter 9 Control via Game Controller

The Thorlabs Kinesis Software includes the possibility of controlling various components
via external input devices. For the Optical Tweezer kit, this allows to drive the KDC101
Servo Motor Controllers and in turn the sample stage movement via a game controller.
Please note that no game controller is provided with the kit. In the following, the required
settings are explained. They have been tested with a Logitech F310 controller under
Windows 10. While the feature could work with similar devices, we do not guarantee this.
1. Connect both KDC101 controllers as well as the game controller to your PC via
USB, then start the Kinesis software.
2. From the main menu of the Kinesis software, choose File -> Input Devices.
3. In the upper right corner of the window that has opened, select your controller
from the dropdown menu.
4. Optionally, you can turn the laser on/off via the game controller. To do this,
connect your KLD101 to the PC via USB, select its serial number in the Target
dropdown menu behind Button A and choose Laser On in the Action dropdown
menu. Do the same for Button B and select Laser Off as Action.
5. On the right side of the same window, select the Target dropdown menu of Left
Analog Y, then choose the serial number of the KDC101 that is responsible for a
movement of the camera image on the screen in vertical direction. Choose the
other KDC101 in the Target dropdown menu of Left Analog X.
6. In the Action dropdown menus of both Left Analog Y and Left Analog X, select
Move Continuous. The settings should now look similar to Figure 67.
7. Click on the Calibrate-Button next to Left Analog Y. In the window that opens,
check the box in front of Use Custom Banding Values (see Figure 68).
8. In the table appearing below, you can now customize the movement speed of the
stage in relation to the displacement of the analog stick. This is done by pairing
a lower boundary of the joystick displacement with a corresponding movement
speed. Such value pairs can be created by clicking the Add button on the right
side of the table. If the joystick displacement is larger than the boundary, the
stage will move at that speed until the displacement of the next higher boundary
is reached.
The velocity is expressed as a percentage of the maximum speed of the sample stage
motors, which is about 2.5 mm/s. We recommend the setup displayed in Figure 68 for both
axes (Note the checked Invert Sense checkbox for the Y-axis). Depending on the controller
and the intended experiment, other values may be more suitable.
Before using the game controller, the KDC101 controllers should be homed. To do this,
press the Home Button for both controllers in Kinesis (see Figure 69). After homing, the

stage will be in one corner of its travel range and should be moved in positive direction for
both axes to center the sample under the objective.
Input Devices Settings in Kinesis Software
Page 82 MTN012639-D02
Recommended Controller Calibration Settings
Homing the KDC101 controllers

Portable Optical Tweezers Kit Chapter 10: Troubleshooting
Chapter 10 Troubleshooting
 When the stage is moved, the sample quickly moves out of focus and loses the
bead.
We have seen this behavior when the screw on the locking plate is tightened too
much.
 The camera image is stuck/frozen.

If the image is stuck (framerate in status bar not counting), close the program.
Take the USB out and plug it back in. Restart the ThorCam software. Enter the
settings and reduce the pixel clock to 8 - 10 MHz, exposure to 100 ms, and restart
the stream from the camera.
 Actuating the knob on the KLD101 does not change the laser diode current.
Make sure you are in the “Const I” mode. Make sure all settings are correct,
especially the anode grounding. If the knob still does not show an effect, unplug
the USB from the PC, turn the power of the KLD101 off and back on again.
 All settings are correct, but the laser diode still doesn’t emit light.
Switch of the KLD101, then remove the laser diode from its mount and reinsert
it (see Section 6.1.1).
 KDC101 controller shows the message “Overload”.
This message means that the motor was driven to the maximum extent. First, try
to switch the KDC101 off and back on again. Then, move the stage back. If this
does not work, you need to remove the motor from the MT1(/M) stage. Loosen
the screw that holds the barrel and remove the motor while holding both parts
of the stage to avoid rapid retraction and potential damage. When the motor
is removed, turn the KDC101 off and back on. Then, actuate the motor towards
the middle position. Afterwards, insert the motor back into the MT1(/M).
 When the stage is actuated, the camera image moves at an angle.
Make sure the camera is mounted parallel to the cage system.
 The optical tweezers don’t trap beads.
o Check your laser diode’s current. It should be about 95 mA.
Page 84 MTN012639-D02
Portable Optical Tweezers Kit Chapter 10: Troubleshooting
o You can try to vary the distance between the lenses in the beam
expander section. 12 cm is the maximum distance (measured from the
lens centers); you should not exceed it. You should also not go below
11 cm.
o Make sure your camera’s plane is set correctly, see Section 6.3.
o Verify that the beam expander cage assembly is parallel to the
breadboard using a level.
o You may try to slightly vary the collimation lens’ distance to the laser
diode.
o Make sure the laser spot hits the objective’s aperture at its center.
o Is the objective clean? Are you able to find the plane with the beads in
their solution? If not, you may need to carefully clean the objective.
o Have you used immersion oil? Please note that this setup does NOT
require immersion oil. The objective is an air objective so please refrain
from using immersion oil.
o Make sure there is enough liquid in the sample, see Figure 48. If there
is not enough liquid, the beads will not be able to move.
o After a while, the beads will stick. You may need to prepare a new
sample, or you can try to move the coverslip slightly to dislodge the
beads (make sure to move the stage away from the objective first to
avoid damage).
o If all of the above points do not show an improvement, please carefully
repeat all steps in the adjustment procedure, Sections 6.4 and 6.5.

Portable Optical Tweezers Kit Chapter 11: Regulatory
Chapter 11 Regulatory
As required by the WEEE (Waste Electrical and Electronic Equipment Directive) of the
European Community and the corresponding national laws, Thorlabs offers all end users
in the EC the possibility to return “end of life” units without incurring disposal charges.
 This offer is valid for Thorlabs electrical and electronic
equipment:
 Sold after August 13, 2005
 Marked correspondingly with the crossed out “wheelie
bin” logo (see right)
 Sold to a company or institute within the EC
 Currently owned by a company or institute within the EC
 Still complete, not disassembled and not contaminated
As the WEEE directive applies to self contained operational electrical and electronic
products, this end of life take back service does not refer to other Thorlabs products, such
as:
 Pure OEM products, that means assemblies to be built into a unit by the user
(e.g. OEM laser driver cards)
 Components
 Mechanics and optics
 Left over parts of units disassembled by the user (PCB’s, housings etc.).
If you wish to return a Thorlabs unit for waste recovery, please contact Thorlabs or your
nearest dealer for further information.
Waste Treatment is Your Own Responsibility
If you do not return an “end of life” unit to Thorlabs, you must hand it to a company
specialized in waste recovery. Do not dispose of the unit in a litter bin or at a public waste
disposal site.
Ecological Background
It is well known that WEEE pollutes the environment by releasing toxic products during
decomposition. The aim of the European RoHS directive is to reduce the content of toxic
substances in electronic products in the future.
The intent of the WEEE directive is to enforce the recycling of WEEE. A controlled recycling
of end of life products will thereby avoid negative impacts on the environment.
Page 86 MTN012639-D02
Portable Optical Tweezers Kit Chapter 12: Thorlabs Worldwide Contacts
Chapter 12 Thorlabs Worldwide Contacts

USA, Canada, and South America UK and Ireland
Thorlabs, Inc. Thorlabs Ltd.
56 Sparta Avenue 1 Saint Thomas Place
Newton, NJ 07860 Ely CB7 4EX
USA Great Britain
Tel: 973-300-3000 Tel: +44 (0) 1353-654440
Fax: 973-300-3600 Fax: +44 (0) 1353-654444
www.thorlabs.com www.thorlabs.com
www.thorlabs.us (West Coast) Email: sales.uk@thorlabs.com
Email: sales@thorlabs.com Support: techsupport.uk@thorlabs.com
Support: techsupport@thorlabs.com
Europe Scandinavia
Thorlabs GmbH Thorlabs Sweden AB
Hans-Böckler-Str. 6 Bergfotsgatan 7
85221 Dachau / Munich 431 35 Mölndal
Germany Sweden
Tel: +49-(0) 8131-5956-0 Tel: +46-31-733-30-00
Fax: +49-(0) 8131-5956-99 Fax: +46-31-703-40-45
www.thorlabs.de www.thorlabs.com
Email: europe@thorlabs.com Email: scandinavia@thorlabs.com
France Brazil
Thorlabs SAS Thorlabs Vendas de Fotônicos Ltda.
109, rue des Côtes Rua Riachuelo, 171
78600 Maisons-Laffitte São Carlos, SP 13560-110
France Brazil
Tel: +33 (0) 970 444 844 Tel: +55-16-3413 7062
Fax: +33 (0) 825 744 800 Fax: +55-16-3413 7064
www.thorlabs.com www.thorlabs.com
Email: sales.fr@thorlabs.com Email: brasil@thorlabs.com
Japan China
Thorlabs Japan, Inc. Thorlabs China
3-6-3, Kitamachi, Room A101, No. 100, Lane 2891, South
Nerima-ku, Tokyo 179-0081 Qilianshan Road
Japan Putuo District
Tel: +81-3-6915-7701 Shanghai 200331
Fax: +81-3-6915-7716 China
www.thorlabs.co.jp Tel: +86 (0) 21-60561122
Email: sales@thorlabs.jp Fax: +86 (0) 21-32513480
www.thorlabschina.cn
Email: chinasales@thorlabs.com

www.thorlabs.com

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EDU-OT2

6.2. Controller and Software Setup ................................................44

6.3. Camera Adjustment ................................................................54

Chapter 7 Experiments ............................................................................... 63

Chapter 8 Teaching Tips ............................................................................ 73

Chapter 1 Warning Symbol Definitions

Both Direct and Alternating Current

Earth Ground Terminal

Protective Conductor Terminal

Frame or Chassis Terminal

In Position of a Bi-Stable Push Control

Out Position of a Bi-Stable Push Control

Caution: Risk of Electric Shock

Caution: Hot Surface

Caution: Risk of Danger

Warning: Laser Radiation

Rev B, July 8, 2019 Page 1

Chapter 3 Product Description

Laser Beam Path

The light path through the optical tweezers experiment.

Chapter 4 Principles of Optical Tweezers

4.1. Dipole Approach in the Rayleigh Scattering Regime R << 

Rev B, July 8, 2019 Page 5

4.2. Geometrical Optics Approach in the Mie Regime R >> 

𝑃𝑟𝑒𝑓𝑙 = 𝑃𝐵𝑒𝑎𝑚 ⋅ 𝑅𝑟 (11)

𝑃𝑡𝑟𝑎𝑛𝑠 = 𝑃𝐵𝑒𝑎𝑚 ⋅ 𝑇 (12)

Reflection and Transmission of an Incident Partial Beam on the

𝑇2 (cos(2𝜃 − 2𝑡) + 𝑅𝑟 cos(2𝜃))

𝑇2 (sin(2𝜃 − 2𝑡) + 𝑅𝑟 cos(2𝜃))

Q Factor Angular Dependence5

In the following, we’ll discuss these forces in more detail.

Total force on the sphere

For that, we integrate (i) over the distance 𝑟

Figure 4 shows how the coordinates 𝑟 and 𝛽

Coordinates of a Partial Beam

𝑋 = sin 𝜙 ∙ 𝑆 = sin 𝜃 ∙ 𝑅 (19)

Relation Between 𝒓 and 𝜽

Rev B, July 8, 2019 Page 11

The Contribution of One Ray to the Total Force5,6

𝐹𝑠,𝑡𝑜𝑡 = ∫ ∫ 𝐹𝑠 ∙ cos 𝜙𝑟𝑑𝑟𝑑𝛽 (25)

𝐹𝑔,𝑡𝑜𝑡 = ∫ ∫ −𝐹𝑔 ∙ sin 𝜙𝑟𝑑𝑟𝑑𝛽 (26)

Gradient force, 𝐹𝑔,𝑡𝑜𝑡

Location of the focus in units of the sphere radius 𝑅 (Z direction)

Rev B, July 8, 2019 Page 13

Gradient force, 𝐹𝑔,𝑡𝑜𝑡

Location of the focus in units of the sphere radius 𝑅 (Y direction)

Scattering and Gradient Force with the Focus Changed in Y

Influence of the Numerical Aperture

Behavior of the Trap’s Strength with Respect to the Numerical

Rev B, July 8, 2019 Page 15

Influence of the Laser Power

Chapter 5 Kit Components

5.1. Trapping Laser Source

Rev B, July 8, 2019 Page 17

5.2. Beam Expander

1 x LA1509-A 1 x SM1A6 1 x SM05L03

5.3. Right-Angle Mirrors

Rev B, July 8, 2019 Page 19

5.4. Sample Positioning System

2 x MT405 1 x Slide Holder Part 1 1 x Slide Holder Part 2

Rev B, July 8, 2019 Page 21

5.6. Additional Components